Immunoexpression of P63 and SOX2 in triple-negative breast cancers, Indonesia

Reno K Kamarlis; Muhammad ND Lubis; Bethy S Hernowo; Azmi S Kar

doi:10.12688/f1000research.12671.2

Home Browse Immunoexpression of P63 and SOX2 in triple-negative breast cancers,...

ALL Metrics

Views

Downloads

Get PDF

Get XML

Export

▬

✚

Research Article

Revised

Immunoexpression of P63 and SOX2 in triple-negative breast cancers, Indonesia

[version 2; peer review: 2 approved]

Reno K Kamarlis^1,2, Muhammad ND Lubis³, Bethy S Hernowo⁴, Azmi S Kar⁵

PUBLISHED 08 Jan 2018

Author details Author details

¹ Department of Pathology Anatomy, Dr. Zainoel Abidin General Hospital, Banda Aceh, Indonesia
² Department of Pathology Anatomy, School of Medicine, Syiah Kuala University, Banda Aceh, Indonesia
³ Department of Pathology Anatomy, Medical Faculty, Sumatera Utara University, Medan, Indonesia
⁴ Department of Pathology Anatomy, Medical Faculty, Padjajaran University, Bandung, Indonesia
⁵ Department of Internal Medicine, Medical Faculty, Sumatera Utara University, Medan, Indonesia

Reno K Kamarlis
Roles: Conceptualization, Data Curation, Formal Analysis, Funding Acquisition, Methodology, Resources, Validation, Visualization

Muhammad ND Lubis
Roles: Conceptualization, Data Curation, Investigation, Methodology, Supervision, Validation

Bethy S Hernowo
Roles: Conceptualization, Data Curation, Funding Acquisition, Investigation, Methodology, Resources, Supervision, Validation

Azmi S Kar
Roles: Conceptualization, Data Curation, Funding Acquisition, Resources, Supervision, Validation

OPEN PEER REVIEW

REVIEWER STATUS

Abstract

Background: Using immunohistochemical stains to target specific breast cancer markers has become indispensable for evaluation of small diagnostic tissue specimens, and therefore novel marker cocktails for specific breast cancers are required. This study was conducted to assess the immunoexpression of P63 and SOX2 in triple negative breast cancer (TNBC), and to evaluate the predictive diagnostic value of these markers for specific types of TNBC.
Methods: Histological slides and paraffin blocks of TNBC cases were collected from Dr. Hasan Sadikin Hospital, Bandung, Indonesia from 5-years period (2011-2015). Each histological slide was subjected to immunohistochemical staining for P63 (nucleus and cytoplasm) and SOX2 (nucleus), with specific primer antibodies. Immunoexpression of P63 and SOX2 was evaluated using immunoreactivity scoring. Associations between P63 and SOX2 immunoexpression and TNBC types were assessed using Mann Whitney tests. In addition, the predictive diagnostic values of these markers were assessed.
Results: Forty TNBC histological slides were included, and 23 (57.5%) were Basal-like type TNBC and 17 (42.5%) were Non basal-like type TNBC. Immunoexpression of P63 nucleus and SOX2 was not different between types of TNBC. However, immunoexpression of P63 in the cytoplasm in Basal-like type TNBC was significantly higher than in Non basal-like type TNBC (p=0.021). Predictor diagnostic value analysis suggested that immunoexpression of P63 in cytoplasm had 56.5% sensitivity and 70.6% specificity for diagnosing Basal-like type TNBC, with area under curve of 0.64.
Conclusions: Immunoexpression of P63 in the cytoplasm has a relatively weak diagnostic value to discriminate Basal-like and Non basal-like types of TNBC.

Keywords

breast cancer, breast cancer marker, P63, SOX2, triple negative breast cancers

Corresponding author: Reno K Kamarlis

Competing interests: No competing interests were disclosed.

Grant information: The author(s) declared that no grants were involved in supporting this work.

Copyright: © 2018 Kamarlis RK et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Data associated with the article are available under the terms of the Creative Commons Zero "No rights reserved" data waiver (CC0 1.0 Public domain dedication).

How to cite: Kamarlis RK, Lubis MN, Hernowo BS and Kar AS. Immunoexpression of P63 and SOX2 in triple-negative breast cancers, Indonesia [version 2; peer review: 2 approved]. F1000Research 2018, 6:1780 (https://doi.org/10.12688/f1000research.12671.2) First published: 28 Sep 2017, 6:1780 (https://doi.org/10.12688/f1000research.12671.1) Latest published: 08 Jan 2018, 6:1780 (https://doi.org/10.12688/f1000research.12671.2)

Revised Amendments from Version 1

We revised the introduction focusing on TNBC epidemiology. We emphasised the introduction related to TNBC classification, characteristics and behaviour. We also updated the role of p63 in cancer within the introduction.
In the conclusion, we included more information related to the frequency of Basal and Non-basal type of TNBC and this finding related to the expression of SOX2 and P63 both in Basal and Non-basal type of TNBC.
We also included the acknowledgement section in the current version. We also revised the author list; Harapan Harapan, MD was requested to be mentioned in the acknowledgement section rather than as co-author due to current changing of his research field of interest.

See the authors' detailed response to the review by Irianiwati Widodo
See the authors' detailed response to the review by Diah Rini Handjari

Introduction

Breast cancer, accounting for 25% of all cancer cases and 15% of all cancer deaths among females, is the most frequently diagnosed cancer among female worldwide^1,2. The incidence of breast cancer increased significantly, approximately by 30% in developed countries³ and currently it has been also rising in many developing countries². In Asia, 639,824 breast cancer cases and 228,926 deaths were recorded in 2012, from which 48,998 cases and 19,750 deaths occurred in Indonesia⁴. Triple-negative breast cancer (TNBC), a group of breast cancers with the absence of oestrogen receptor and progesterone receptor and no overexpression of human epidermal growth factor receptor 2 (HER2), represents 10%–20% of invasive breast cancer. A global data base, National Cancer Data Base (NCDB), reveals that TNBC was present in 13% of breast cancer patients, ranged from 23.7% in African-Americans to 8.9% in Filipino patients⁵. In Southeast Asia, a study found that TNBC presented in 10.5% among 1227 breast cancer patients⁶ . In Indonesia, a study that was conducted between 2010 and 2011 in Bandung found that 11.9% of breast cancer patients were TNBC⁷.

Immunohistochemically, TNBC could be divided into two subtypes: Basal-like (positive for the expression of high-molecular-weight/basal cytokeratins 5/6 (CK5/6) and epidermal growth factor receptor (EGFR)) and Non basal-like (negative for the expression of CK5/6 and EGFR). Basal-like TNBC usually has p53 mutation, EGFR overexpression, loss of function of BRCA1, c-MYC amplification, and high histological grade indicating more aggressive characteristics and aggressive behavior^8,9. In addition, majority of Basal-like cancer cannot be managed effectively with trastuzumab and hormonal treatments¹⁰.

Advanced screening and diagnosis methods for breast cancer such as mammograms, ultrasound, magnetic resonance imaging and fine-needle aspiration, have allowed for detection of small lesions at the early stage. Identifying breast cancer at the early stage will increase the potential for curative treatment and therefore increases the survival rate^11–14. However, smaller lesions are more challenging to diagnose. Therefore, it is essential to use an advanced immunohistochemical approach for evaluation of smaller tissue specimens that target more specific markers¹⁵.

A previous study using a MCF7 breast cancer cell line to produce MCF7-derived tumour xenografts found that P63 and SOX2 immunostainings were two potential markers for breast cancer¹⁶. P63, involved in cellular differentiation, is a homolog of tumour protein P53 and in normal breast ducts and lobules it is expressed frequently in the nuclei of myoepithelial cells¹⁷. Mutation of the p53 gene results in a very high risk of breast cancer¹⁸. The roles of p63 in tumorigenesis, cancer progression, and metastasis are still being discovered. However, in animal model found that p63 deficiency may be a causative factor for metastatic spread^19,20. In addition, clinical evidence suggests that a robust correlation between reduced p63 expression and cancer progression²¹.

A study revealed that the total percentage of P63-positive cells was related to marked nuclear pleomorphism and the intensity of P63 staining was associated with syncytial growth pattern in TNBC²². In addition, data also reveals that p63 gene expression in breast cancer could be used as a specific marker of metaplastic carcinoma¹⁷, and P63 immunohistochemical staining could improve diagnostic accuracy of breast cancer even in small tissue specimens²³.

SOX2 is a transcription factor belonging to the SOX family and functions as an activator or suppressor of gene transcription^24,25. Data shows that SOX2 promotes cellular proliferation of breast tissue²⁶ and regulates self-renewal in cancer stem cells²⁷. The scientific evidence reveals that SOX2 acts as an oncogene in epithelial cancers²⁵ and in the breast, a study found that silencing of sox2 gene was associated with reduction of the size of the cancer stem cells and restoration of tamoxifen sensitivity²⁸. All together, these data indicate that P63 and SOX2 have pivotal role in breast cancer and therefore are potential to be used as specific biomarkers. This study was conducted to assess the immunoexpression of P63 and SOX2 in TNBC cases in order to provide insight regarding their potential diagnostic value (single or in combination) to differentiate TNBC types.

Methods

Study setting and histological slides

A cross-sectional study to assess the immunoexpression of P63 and SOX2 in TNBC cases (negative expression of estrogen and progesterone receptors and c-erbB2) was conducted. Histological slides of TNBC and their paraffin blocks, tested between the 1^st of January 2011 and 31^st of December 2015, were collected from the Pathology Anatomy Laboratory, Dr. Hasan Sadikin Hospital, Bandung, Indonesia. Each histological slide was examined by two certified pathologists. To classify the type of TNBC morphology, between Basal-like type TNBC and Non basal-like type TNBC, cytokeratin 5/6 (CK 5/6) immunohistochemical staining was carried out on all TNBC histological slides. Concurrently, the immunoexpression of P63 and SOX2 was measured using immunohistochemical stains with specific primer antibodies. The protocol of this study was approved by the Health Research Ethical committee of Sumatera Utara University (approval 103/KOMET/FK USU/2015) and the usage of histological specimens was approved by the Pathology Anatomy Laboratory of Dr. Hasan Sadikin Hospital (LB.02.01/B29/239/X/2015).

Immunohistochemistry

Forty archival paraffin blocks from TNBC cases were subjected to immunohistochemical staining to assess the immunoexpression of CK 5/6, P63 and SOX2. Briefly, 4 μm sections of each paraffin block were prepared using standard procedure²⁹. Immunohistochemical staining was conducted using primary antibodies as follows: anti-CK5/6 monoclonal antibody (Biocare Medical, Concord, CA, USA), anti-P63 monoclonal antibody (Biocare Medical, Concord, CA, USA) and anti-SOX2 monoclonal antibody (Abcam, Cambridge, UK). Starr Trek Universal HRP Detection (Biocare Medical, Concord, CA, USA) was used as second antibody. A chromogen 3,3’-diaminobenzidine (DAB) (Biocare Medical, Concord, CA, USA) was used to develop the colour. For each experiment, appropriate controls were used.

Immunoexpression of CK 5/6 was interpreted as positive or negative, in which positive CK 5/6 indicates Basal-like type TNBC while negative CK 5/6 indicates Non basal-like type TNBC. Immunoexpression of P63 and SOX2 was evaluated using an immunoreactivity scoring system that had been published elsewhere with modification²². Staining intensity was scored as follows: 1 (no staining), 2 (weak staining), 3 (moderate staining) and 4 (strong staining). The percentage of positively stained tumour cells was assessed as a proportion of the total number of tumour cells present in the section as follows: 1 (<20%), 2 (≥20–50%), 3 (>50–80%) and 4 (>80%).

Immunoreactivity score was calculated by multiplying staining intensity and the percentage of positivity, and the score therefore ranged from 1 to 16. The immunoreactive score was then divided into low (≤ 5), moderate (≥ 6 – 10) and high (≥11 – 16). Immunoexpression of P63 was measured both in cytoplasm and nucleus while SOX2 immunoexpression was measured in nucleus only.

Statistical analysis

Normality of the data was assessed using the Shapiro-Wilk test and therefore the analysis tests chosen based on the normality of the data. The correlations between immunoexpression of P63 (cytoplasm and nucleus) and SOX2 were assessed using Pearson correlation and Spearman correlation, respectively. The associations of P63 and SOX2 immunoexpression and type of TNBC were assessed using Mann Whitney test. The predictive diagnostic values of P63 cytoplasm for diagnosing Basal-like type TNBC were estimated using several immunoreactivity score cut-off points. Receiver operating characteristic curve (ROC) was plotted and area under the ROC curves (AUC) was estimated. For all analyses, estimates were considered statistically significant for two-tailed values of p<0.05. All analyses were conducted using Statistical Package for the Social Sciences software (SPSS for Windows, Version 16, Chicago, IL).

Results

Clinicopathology and classification of TNBC

The histopathology of the TNBC samples used in this study is described in Table 1. Approximately 45% of the samples were classified as metaplastic carcinomas. In addition, immunohistochemical staining for CK 5/6 revealed that 23 (57.5%) of samples were Basal-like type TNBC and while 17 (42.5%) samples were Non basal-like type TNBC.

Table 1. Histopathology types of TNBC samples used in this study (N=40).

Type of histopathology	n (%)
Metaplastic carcinoma, spindle cells component	10 (25.0)
Metaplastic carcinoma, producing mucin	3 (7.5)
Metaplastic carcinoma, liposarcoma component	2 (5.0)
Metaplastic carcinoma, squamous cell	1 (2.5)
Metaplastic carcinoma, matrix hyaline	1 (2.5)
Medullary carcinoma	13 (32.5)
Micropapillary carcinoma	1 (2.5)
Invasive ductal carcinoma grade 3	6 (15.0)
Invasive lobular carcinoma grade 3	2 (5.0)
Invasive ductal carcinoma + invasive lobular carcinoma (mixed)	1 (2.5)

Immunoreactivity score of P63 and SOX2

Immunoexpression of P63 and SOX2 in samples, categorized by immunoreactivity score, are presented in Table 2. For both types of TNBC (basal and non basal-like type), all immunoreactivity scores for P63 in the nucleus were classified as low grade, while 11 (27.5%) and 7 (17.5%) samples were classified as moderate and high grade, respectively for the P63 in the cytoplasm. The immunoreactivity grade for SOX2 was similar to P63 in the cytoplasm, and therefore correlation analyses were conducted.

Table 2. Immunoreactivity score of P63 and SOX2 in TNBC.

Grade	P63 cytoplasm, n (%)	P63 nucleus, n (%)	SOX2, n (%)
Low (≤ 5)	22 (55.0)	40 (100.0)	19 (47.5)
Moderate (≥ 6 – 10)	11 (27.5)	0 (0.0)	12 (30.0)
High (≥11 – 16)	7 (17.5)	0 (0.0)	9 (22.5)

Correlation between immunoexpression of P63 and SOX2

There was a strong negative correlation between immunoexpression of P63 in the cytoplasm and immunoexpression of SOX2 in the nucleus in metaplastic carcinoma (a sub-type of TNBC basal-like type) (r=-0.73, p=0.013) (Table 3). In addition, linear regression showed a relatively strong correlation between P63 cytoplasm and SOX2 immunoexpression in metaplastic carcinoma (r=0.49, p=0.012). There was no significant correlation between P63 cytoplasm and SOX2 immunoexpression in Non basal-like type TNBC, and no significant correlation between P63 nucleus and SOX2 immunoexpression either in Basal-like type or Non basal-like type of TNBC.

Table 3. Correlation between immunoexpression of P63 cytoplasm and SOX2 in Basal-like type TNBC.

TNBC basal-like type	n	P63	SOX2	r	p
TNBC basal-like type	n	Mean (±SD)	Mean (±SD)	r	p
Invasive ductal carcinoma	2	5.50 (3,54)	5.00 (5.66)	-	-
Invasive ductal carcinoma and invasive lobular carcinoma	1	16.00 (0.00)	9.00 (0.00)	-	-
Invasive lobular carcinoma	1	8.00 (0.00)	1.00 (0.00)	-	-
Invasive micropapillary carcinoma	1	12.00 (0.00)	16.00 (0.00)	-	-
Medullary carcinoma	6	5.50 (4.76)	8.00 (5.48)	0.64	0.172
Metaplastic carcinoma	12	6.67 (4.68)	6.00 (3.77)	-0.73	0.013*

*Significant at 0.05

Immunoexpression of P63 and SOX2 in Basal-like and Non basal-like type of TNBC

Immunoexpression of P63 cytoplasm, P63 nucleus and SOX2 in Basal-like and Non Basal-like TNBC is shown in Table 4. The data indicates that the immunoexpression of P63 cytoplasm in Basal-like type TNBC was significantly higher compared to Non basal-like type TNBC (p=0.021). Immunoexpression of P63 nucleus and SOX2 was not different between Basal-like and Non basal-like types of TNBC, with p-values of p=0.27 and p=0.17, respectively.

Table 4. Immunoexpression of P63 and SOX2 in Basal-like type TNBC and Non basal-like type TNBC.

Marker	TNBC type	n	Immunoreactivity score Mean (±SD)	p
P63 cytoplasm	Basal-like	23	6.96 (4,73)	0.021*
P63 cytoplasm	Non basal-like	17	3.76 (4,16)	0.021*
P63 nucleus	Basal-like	23	1.22 (0,52)	0.273
P63 nucleus	Non basal-like	17	1.06 (0,24)	0.273
SOX2	Basal-like	23	6.78 (4,69)	0.172
SOX2	Non basal-like	17	4.82 (3,61)	0.172

*Significant at 0.05

Predictor diagnostic value of P63 in the cytoplasm for diagnosing Basal like type TNBC

As mentioned above, immunoexpression of P63 in the cytoplasm was the only marker that was significantly different between TNBC types. Therefore, immunoreactivity score of P63 cytoplasm was further analysed to determine its ability to predict Basal-like type TNBC. Table 5 shows the predictive values of P63 in the cytoplasm for determining Basal-like type TNBC, using seven immunoreactivity score cut-off values from 3 to 9. It shows that P63 has relatively weak diagnostic value in diagnosing Basal-like type TNBC. The highest sensitivity was achieved at immunoreactivity score 3, while specificity was increasing with a higher immunoreactivity score.

Table 5. Predictor diagnostic value of P63 in the cytoplasm for diagnosing Basal-like type TNBC.

Diagnostic test	Cut-off of P63 cytoplasm immunoreactivity score
Diagnostic test	3	4	5	6	7	8	9
Sensitivity (%)	78.3	73.9	56.5	56.5	52.2	52.2	30.4
Specificity (%)	58.8	58.8	70.6	70.6	82.4	82.4	88.2
Positive predictive value (%)	72.0	70.8	72.2	72.2	80.0	80.0	77.8
Negative predictive value (%)	66.7	62.5	54.5	54.5	56.0	56.0	48.4
Positive likelihood ratio (%)	190.1	179.5	192.2	192.2	295.7	295.7	258.7
Negative likelihood ratio (%)	37.0	44.3	61.6	61.6	58.1	58.1	78.8
Accuracy (%)	70.0	67.5	62.5	62.5	65.0	65.0	55.0

Using the average score of P63 cytoplasm immunoexpression for Basal-like type TNBC in this study, 5.6 or 6, the sensitivity and specificity of P63 cytoplasm immunoreactivity score to predict Basal-like type TNBC was 56.5% and 72.6%, respectively with area under curve 0.64. The receiver operating curve of predictive diagnostic value of P63 cytoplasm for determining Basal-like type TNBC is plotted in Figure 1.

Figure 1. Receiver operating curve of P63 cytoplasm immunoexpression for determining TNBC basal-like type.

Dataset 1.Immunoexpression and immunoreactivity scores of P63, SOX2 and CK 5/6 in the forty specimens that were analysed.

Discussion

To the best of our knowledge, this is the first study conducted to assess the immunoexpression of P63 and SOX2 in TNBC cases in Indonesia. Some studies have been conducted to assess the predictive values of P63 as specific marker for breast cancer^17,30. In addition, the idea of utilization of a cocktail of specific markers has been proposed previously to provide higher sensitivity and specificity for diagnosing specific breast cancers^15,22,31. However, none of the previous studies had been conducted to assess the diagnostic value of immunoexpression of P63 and SOX2 in combination. This study, at the beginning, sought to assess predictive value of combination both of those markers for specific type of TNBC. However, we found that there was no difference in immunoexpression of SOX2 between Basal-like type TNBC and Non basal-like type TNBC. Nevertheless, we found that immunoexpression of P63 cytoplasm, but not P63 nucleus, was higher in Basal-like type TNBC compared to Non basal-like type TNBC.

P63 has been proposed as a breast cancer marker for a long time, but with conflicting results. A study demonstrated that immunoexpression of P63 was associated with breast cancers, for example the metaplastic carcinoma type of breast cancer¹⁷, but there was no difference in immunoexpression of P63 between medullary breast carcinomas and atypical medullary breast carcinomas of TNBC³⁰. In our study, we found that the sensitivity and specificity of P63 cytoplasm immunoexpression to diagnose Basal-like type TNBC was 56.5% and 72.6%, respectively, with area under curve of 0.64. This sensitivity and specificity seems higher compared to a previous study, with 14% and 94%, respectively in determining a Basal-like type in infiltrative ductal carcinomas (TNBC)²². All together, these data indicate a weak predictive value of P63 immunoexpression as marker for Basal-like type TNBC. However, a study found that P63 is a specific marker for metaplastic carcinomas of the breast (a sub-type of Basal-like type TNBC)¹⁷. In our study, we could not assess the predictive value of P63 cytoplasm immunoexpression for determining metaplastic carcinomas, due to the small sample size (see Table 3).

We found that SOX2 immunoexpression grade was classified as moderate and high grade in 55% of TNBC cases (Table 2,) and it has been indicated previously that SOX2 has strong roles in promoting breast cancers^26–28,32. However, there was no different in immunoexpression between Basal-like type TNBC and Non basal-like type TNBC. This result indicates that SOX2 expression is not different amongst TNBC types. This finding was in line with a previous study that indicated that SOX2 was expressed across different breast cancer subtypes³³. A study found that SOX2 antibody in the sera was is higher in patients with breast cancer compared to healthy women and therefore it could be used to discriminate between breast cancer patients and healthy controls³⁴. In addition, a meta-analysis found that SOX2 expression was associated with tumor size, histological grade, the aggressiveness and lymph node metastasis in TNBC patients³⁵. All together, these results indicate that there was a possibility SOX2 expression could be used for diagnosing breast cancers, but there was no difference in expression amongst breast cancer types, and therefore it could not be used as specific marker for differentiating TNBC types.

There are some limitations to this study. The sample size was relatively small, and therefore some analyses could not be conducted. In addition, the diagnostic specimens were collected from different procedures such as from biopsy, mastectomy or lumpectomy, and this might influence the immunoexpression of the markers.

Conclusions

Immunoexpression of P63 cytoplasm is higher among Basal-like type TNBC compared to Non basal-like type TNBC. However, the predictive diagnostic value of P63 immunoexpression in the cytoplasm for Basal-like type TNBC is relatively low, with 56.5% sensitivity and 72.6% specificity.

Data availability

Dataset 1: Immunoexpression and immunoreactivity scores of P63, SOX2 and CK 5/6 in the forty specimens that were analysed. DOI, 10.5256/f1000research.12671.d179131³⁶

Competing interests

No competing interests were disclosed.

Grant information

The author(s) declared that no grants were involved in supporting this work.

Acknowledgments

We would like to thank Harapan Harapan, MD (https://orcid.org/0000-0001-7630-8413) from Medical Research Unit, School of Medicine, Syiah Kuala University, who assisted in revising the manuscript and provided intellectual contribution to this study.

Faculty Opinions recommended

References

1. Key TJ, Verkasalo PK, Banks E: Epidemiology of breast cancer. Lancet Oncol. 2001; 2(3): 133–140. PubMed Abstract | Publisher Full Text
2. Torre LA, Bray F, Siegel RL, et al.: Global cancer statistics, 2012. CA Cancer J Clin. 2015; 65(2): 87–108. PubMed Abstract | Publisher Full Text
3. Althuis MD, Dozier JM, Anderson WF, et al.: Global trends in breast cancer incidence and mortality 1973–1997. Int J Epidemiol. 2005; 34(2): 405–412. PubMed Abstract | Publisher Full Text
4. Ghoncheh M, Momenimovahed Z, Salehiniya H: Epidemiology, Incidence and Mortality of Breast Cancer in Asia. Asian Pac J Cancer Prev. 2016; 17(S3): 47–52. PubMed Abstract | Publisher Full Text
5. Plasilova ML, Hayse B, Killelea BK, et al.: Features of triple-negative breast cancer: Analysis of 38,813 cases from the national cancer database. Medicine (Baltimore). 2016; 95(35): e4614. PubMed Abstract | Publisher Full Text | Free Full Text
6. Alcantara VS, Lim GH, Lim SH, et al.: Incidence and prognosis of non-metastatic triple negative breast cancer (TNBC) among different races in Southeast Asia. J Surg Oncol. 2017; 115(5): 523–537. PubMed Abstract | Publisher Full Text
7. Kusumadjayanti N, Badudu DF, Hernowo BS: Characteristics of patients with estrogen receptor (ER)-negative, progesterone receptor (PR)-negative, and HER2-negative invasive breast cancer in Dr. Hasan Sadikin General Hospital, Bandung, Indonesia from 2010 to 2011. Althea Med J. 2015; 2(3): 391–394. Publisher Full Text
8. Cleator S, Heller W, Coombes RC: Triple-negative breast cancer: therapeutic options. Lancet Oncol. 2007; 8(3): 235–44. PubMed Abstract | Publisher Full Text
9. Kobayashi S: Basal-like subtype of breast cancer: a review of its unique characteristics and their clinical significance. Breast Cancer. 2008; 15(2): 153–8. PubMed Abstract | Publisher Full Text
10. Rakha EA, Reis-Filho JS, Ellis IO: Basal-like breast cancer: a critical review. J Clin Oncol. 2008; 26(15): 2568–2581. PubMed Abstract | Publisher Full Text
11. AIHW: Breast cancer survival by size and nodal status in Australia. In: Registries NBCCAAoC, ed. Cancer series no. 39. Canberra: AIHW; 2007. Reference Source
12. Allemani C, Minicozzi P, Berrino F, et al.: Predictions of survival up to 10 years after diagnosis for European women with breast cancer in 2000–2002. Int J Cancer. 2013; 132(10): 2404–2412. PubMed Abstract | Publisher Full Text
13. Allemani C, Weir HK, Carreira H, et al.: Global surveillance of cancer survival 1995–2009: analysis of individual data for 25,676,887 patients from 279 population-based registries in 67 countries (CONCORD-2). Lancet. 2015; 385(9972): 977–1010. PubMed Abstract | Publisher Full Text | Free Full Text
14. Naroda S, Iqbala J, Miller AB: Why have breast cancer mortality rates declined? J Cancer Policy. 2015; 5: 8–17. Publisher Full Text
15. Reisenbichler ES, Ross JR, Hameed O: The clinical use of a P63/cytokeratin7/18/cytokeratin5/14 antibody cocktail in diagnostic breast pathology. Ann Diagn Pathol. 2014; 18(6): 313–318. PubMed Abstract | Publisher Full Text
16. Liu Y, Coates PJ, Nenutil R, et al.: Lack of correlation between markers of breast cancer initiating cells. Breast Cancer Res. 2010; 12(Suppl 1): P38. Publisher Full Text | Free Full Text
17. Koker MM, Kleer CG: p63 expression in breast cancer: a highly sensitive and specific marker of metaplastic carcinoma. Am J Surg Pathol. 2004; 28(11): 1506–1512. PubMed Abstract | Publisher Full Text
18. Assi HA, Khoury KE, Dbouk H, et al.: Epidemiology and prognosis of breast cancer in young women. J Thorac Dis. 2013; 5 Suppl 1: S2–8. PubMed Abstract | Publisher Full Text | Free Full Text
19. Flores ER, Sengupta S, Miller JB, et al.: Tumor predisposition in mice mutant for p63 and p73: evidence for broader tumor suppressor functions for the p53 family. Cancer Cell. 2005; 7(4): 363–373. PubMed Abstract | Publisher Full Text
20. Su X, Chakravarti D, Cho MS, et al.: TAp63 suppresses metastasis through coordinate regulation of Dicer and miRNAs. Nature. 2010; 467(7318): 986–990. PubMed Abstract | Publisher Full Text | Free Full Text
21. Bergholz J, Xiao ZX: Role of p63 in Development, Tumorigenesis and Cancer Progression. Cancer Microenviron. 2012; 5(3): 311–322. PubMed Abstract | Publisher Full Text | Free Full Text
22. Thike AA, Cheok PY, Jara-Lazaro AR, et al.: Triple-negative breast cancer: clinicopathological characteristics and relationship with basal-like breast cancer. Mod Pathol. 2010; 23(1): 123–133. PubMed Abstract | Publisher Full Text
23. Harton AM, Wang HH, Schnitt SJ, et al.: p63 Immunocytochemistry improves accuracy of diagnosis with fine-needle aspiration of the breast. Am J Clin Pathol. 2007; 128(1): 80–85. PubMed Abstract | Publisher Full Text
24. Weina K, Utikal J: SOX2 and cancer: current research and its implications in the clinic. Clin Transl Med. 2014; 3: 19. PubMed Abstract | Publisher Full Text | Free Full Text
25. Sarkar A, Hochedlinger K: The sox family of transcription factors: versatile regulators of stem and progenitor cell fate. Cell Stem Cell. 2013; 12(1): 15–30. PubMed Abstract | Publisher Full Text | Free Full Text
26. Stolzenburg S, Rots MG, Beltran AS, et al.: Targeted silencing of the oncogenic transcription factor SOX2 in breast cancer. Nucleic Acids Res. 2012; 40(14): 6725–6740. PubMed Abstract | Publisher Full Text | Free Full Text
27. Leis O, Eguiara A, Lopez-Arribillaga E, et al.: Sox2 expression in breast tumours and activation in breast cancer stem cells. Oncogene. 2012; 31(11): 1354–1365. PubMed Abstract | Publisher Full Text
28. Piva M, Domenici G, Iriondo O, et al.: Sox2 promotes tamoxifen resistance in breast cancer cells. EMBO Mol Med. 2014; 6(1): 66–79. PubMed Abstract | Publisher Full Text | Free Full Text
29. Anderson G, Bancroft J: Tissue processing and microtomy. In: Bancroft JG, M., ed. Theory and practiceal of histological techniques 5th Edition. Edinburgh Churchill Livingstone; 2002; 109–123.
30. Matkovic B, Juretic A, Separovic V, et al.: Immunohistochemical analysis of ER, PR, HER-2, CK 5/6, p63 and EGFR antigen expression in medullary breast cancer. Tumori. 2008; 94(6): 838–844. PubMed Abstract
31. Tacha DE, Bloom K, Kyshtoobayava A, et al.: A double immunostaining technique with a cocktail CK5, CK14, p63, CK7 and CK18 distinguishes between hyperplasia of the usual type, atypical hyperplasia, microinvasive and basal phenotype breast cancers. Modern Pathology. 2009; 22: 388a.
32. Liu K, Xie F, Gao A, et al.: SOX2 regulates multiple malignant processes of breast cancer development through the SOX2/miR-181a-5p, miR-30e-5p/TUSC3 axis. Mol Cancer. 2017; 16(1): 62. PubMed Abstract | Publisher Full Text | Free Full Text
33. Lengerke C, Fehm T, Kurth R, et al.: Expression of the embryonic stem cell marker SOX2 in early-stage breast carcinoma. BMC Cancer. 2011; 11: 42. PubMed Abstract | Publisher Full Text | Free Full Text
34. Sun Y, Zhang R, Wang MJ, et al.: SOX2 autoantibodies as noninvasive serum biomarker for breast carcinoma. Cancer Epidemiol Biomarkers Prev. 2012; 21(11): 2043–2047. PubMed Abstract | Publisher Full Text
35. Zheng Y, Qin B, Li F, et al.: Clinicopathological significance of Sox2 expression in patients with breast cancer: a meta-analysis. Int J Clin Exp Med. 2015; 8(12): 22382–22392. PubMed Abstract | Free Full Text
36. Reno KK, Muhammad NDL, et al.: Dataset 1 in: Immunoexpression of P63 and SOX2 in triple-negative breast cancers, Indonesia. F1000Research. 2017. Data Source

Comments on this article Comments (0)

Version 2

VERSION 2 PUBLISHED 28 Sep 2017

Author details Author details

Reno K Kamarlis
Roles: Conceptualization, Data Curation, Formal Analysis, Funding Acquisition, Methodology, Resources, Validation, Visualization

Muhammad ND Lubis
Roles: Conceptualization, Data Curation, Investigation, Methodology, Supervision, Validation

Bethy S Hernowo
Roles: Conceptualization, Data Curation, Funding Acquisition, Investigation, Methodology, Resources, Supervision, Validation

Azmi S Kar
Roles: Conceptualization, Data Curation, Funding Acquisition, Resources, Supervision, Validation

Competing interests

No competing interests were disclosed.

Grant information

The author(s) declared that no grants were involved in supporting this work.

Article Versions (2)

version 2

Revised

Published: 08 Jan 2018, 6:1780

https://doi.org/10.12688/f1000research.12671.2

version 1

Published: 28 Sep 2017, 6:1780

https://doi.org/10.12688/f1000research.12671.1

© 2018 Kamarlis RK et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Data associated with the article are available under the terms of the Creative Commons Zero "No rights reserved" data waiver (CC0 1.0 Public domain dedication).

Download

Export To

metrics

	Views	Downloads
F1000Research	-	-
PubMed Central Data from PMC are received and updated monthly.	-	-

Citations

SEE MORE DETAILS

CITE

how to cite this article

Kamarlis RK, Lubis MN, Hernowo BS and Kar AS. Immunoexpression of P63 and SOX2 in triple-negative breast cancers, Indonesia [version 2; peer review: 2 approved]. F1000Research 2018, 6:1780 (https://doi.org/10.12688/f1000research.12671.2)

NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article.

track

receive updates on this article

Track an article to receive email alerts on any updates to this article.

Open Peer Review

Version 2

VERSION 2

PUBLISHED 08 Jan 2018

Revised

Views

Reviewer Report 20 Feb 2018

Diah Rini Handjari, University of Indonesia, Jakarta, Indonesia

Approved

https://doi.org/10.5256/f1000research.14420.r29568

This revised version is ... Continue reading

CITE

Report a concern

Respond or Comment

Views

Reviewer Report 06 Feb 2018

Irianiwati Widodo, Department of Anatomical Pathology, Faculty of Medicine, Gadjah Mada University, Yogyakarta, Indonesia

Approved

https://doi.org/10.5256/f1000research.14420.r29569

This revised version is good ... Continue reading

CITE

Report a concern

Respond or Comment

Version 1

VERSION 1

PUBLISHED 28 Sep 2017

Views

Reviewer Report 20 Oct 2017

Diah Rini Handjari, University of Indonesia, Jakarta, Indonesia

Approved with Reservations

https://doi.org/10.5256/f1000research.13722.r26782

I. - In introduction there is only little information regarding of nuclear expression of p63.
   - It doesn't mention about cytoplasmic p63 expression.
     What is significance function of p63 in tumor progression or disease progression?
     It written in result and analysis.
   - In introduction you haven't discussed yet about the incidence of triple negative breast cancer in the worldwide, Asia or Indonesia ?

II. In the methods, to classify TNBC morphology into Basal like and non Basal like, based on only, antibody (CK 5/6) but you didn't perform EGFR staining. Which is one of the marker of TNBC.
    - Is there any reference about the scoring of intensity and the percentage of positivity of expression of p63?
    - Is there any reference about imunoreactive score?

III. In discussion, there is also no information about cytoplasmic p63 and its role in disease progression.
    - Researcher only stated that cytoplasmic p63 expression has predictive value to classify breast cancer into TNBC and there is no comparison of p63 expression sensitivity and specificity with other TNBC marker as like CK 5/6 and EGFR which has been routinely used.

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Yes
Are sufficient details of methods and analysis provided to allow replication by others?

Yes
If applicable, is the statistical analysis and its interpretation appropriate?

I cannot comment. A qualified statistician is required.
Are all the source data underlying the results available to ensure full reproducibility?

Partly
Are the conclusions drawn adequately supported by the results?

Partly

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Pathologist with major interest in colorectal cancer

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above.

CITE

Report a concern

Author Response 08 Jan 2018

Harapan Harapan, Medical Research Unit, School of Medicine, Syiah Kuala University, Banda Aceh, Indonesia

08 Jan 2018

Author Response

Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. We would like to ... Continue reading Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. We would like to confirm that predictive diagnostic analysis in this study was conducted using qualified statistician. There is no evidence to support the importance of the negative correlation between immunoexpression of P63 cytoplasm and immunoexpression SOX2 nucleus in metaplastic carcinoma, and therefore we are unable to discuss this finding in depth. In conclusion section, we have added some additional principal finding as suggested by the reviewer.
Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. We would like to confirm that predictive diagnostic analysis in this study was conducted using qualified statistician. There is no evidence to support the importance of the negative correlation between immunoexpression of P63 cytoplasm and immunoexpression SOX2 nucleus in metaplastic carcinoma, and therefore we are unable to discuss this finding in depth. In conclusion section, we have added some additional principal finding as suggested by the reviewer.
Competing Interests: There is no competing interest in this study. Close
Report a concern
Author Response 08 Jan 2018

Harapan Harapan, Medical Research Unit, School of Medicine, Syiah Kuala University, Banda Aceh, Indonesia

08 Jan 2018

Author Response

Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. In the revised manuscript, ... Continue reading Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. In the revised manuscript, the data of the incidence from global, Southeast Asian countries and Indonesia are included.

In our study, we used the histological paraffin blocks that have been confirmed as TNBC previously by testing ER, PR and HER2. To differentiate Basal like and non Basal like, either CK 5/6 or EGFR staining could be used with no significant different sensitivity and specificity (Livasy et al., 2006; Nielsen et al., 2004). In this study, CK 5/6 was employed to differentiate TNB into Basal like and non Basal like and this procedure was conducted during the study. This method is established method to define basal phenotype (Sasa et al., 2008; Rakha et al., 2007).

In our study, we used the histological paraffin blocks that have been confirmed as TNBC previously by testing ER, PR and HER2. To differentiate Basal like and non Basal like, either CK 5/6 or EGFR staining could be used with no significant different sensitivity and specificity (Livasy et al., 2006; Nielsen et al., 2004). In this study, CK 5/6 was employed to differentiate TNB into Basal like and non Basal like and this procedure was conducted during the study. This method is established method to define basal phenotype (Sasa et al., 2008; Rakha et al., 2007). CK 5/6 and EGFR.

As mentioned Immunohistochemistry Section, the principle of the scoring system for immunoexpression of P63 and SOX2 used in this study was have been elsewhere with modification (Thike et al., 2010). In detailed: Staining intensity was scored as follows: 1 (no staining), 2 (weak staining), 3 (moderate staining) and 4 (strong staining). The percentage of positively stained tumour cells was assessed as a proportion of the total number of tumour cells present in the section as follows: 1 (<20%), 2 (≥20–50%), 3 (>50–80%) and 4 (>80%). Then from staining intensity and percentage of positively stained cells, we created immunoreactivity score by multiplying staining intensity and the percentage of positivity. The score, therefore, ranged from 1 to 16 (divided into low (≤ 5), moderate (≥ 6 – 10) and high (≥11 – 16). However, specifications for staining intensity and the percentage of positivity used in this study are the standard system used by Pathology Anatomy Laboratory of Dr. Hasan Sadikin Hospital Bandung since 1990. These score systems are also adopted in some breast cancer diagnostic centres in Indonesia.

Immunoreactive score system used in this study have been published elsewhere (Thike et al., 2010). We mentioned this in Immunohistochemistry Section of our Methods.
In this study produce all raw data related to: type of TNBC (Basal like and Non Basal-like), subtype of cancer, expression of SOX2, P63 and CK 5/6 including staining intensity, distribution of the positively cells and immunoreactive score.

Reference:
Thike AA, Cheok PY, Jara-Lazaro AR, et al. Triple-negative breast cancer: clinicopathological characteristics and relationship with basal-like breast cancer. Mod Pathol. 2010;23(1):123–133.
Livasy CA, Karaca G, Nanda R, et al. Phenotypic evaluation of the basal-like subtype of invasive breast carcinoma. Mod Pathol 2006;19:264-271
Nielsen TO, Hsu FD, Jensen K, et al. Immunohistochemical and clinical characterization of the basal-like subtype of invasive breast carcinoma Clin Cancer Res, 2004;10:5367-5374
Rakha EA, El-Sayed ME, Green AR, et al. Breast carcinoma with basal differentiation: a proposal for pathology definition based on basal cytokeratin expression. Histopathology. 2007 Mar;50(4):434-8.
Sasa M, Bando Y, Takahashi M, et al. Screening for basal marker expression is necessary for decision of therapeutic strategy for triple-negative breast cancer. J Surg Oncol, 2008; 97: 30-34
Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. In the revised manuscript, the data of the incidence from global, Southeast Asian countries and Indonesia are included.

In our study, we used the histological paraffin blocks that have been confirmed as TNBC previously by testing ER, PR and HER2. To differentiate Basal like and non Basal like, either CK 5/6 or EGFR staining could be used with no significant different sensitivity and specificity (Livasy et al., 2006; Nielsen et al., 2004). In this study, CK 5/6 was employed to differentiate TNB into Basal like and non Basal like and this procedure was conducted during the study. This method is established method to define basal phenotype (Sasa et al., 2008; Rakha et al., 2007).

In our study, we used the histological paraffin blocks that have been confirmed as TNBC previously by testing ER, PR and HER2. To differentiate Basal like and non Basal like, either CK 5/6 or EGFR staining could be used with no significant different sensitivity and specificity (Livasy et al., 2006; Nielsen et al., 2004). In this study, CK 5/6 was employed to differentiate TNB into Basal like and non Basal like and this procedure was conducted during the study. This method is established method to define basal phenotype (Sasa et al., 2008; Rakha et al., 2007). CK 5/6 and EGFR.

As mentioned Immunohistochemistry Section, the principle of the scoring system for immunoexpression of P63 and SOX2 used in this study was have been elsewhere with modification (Thike et al., 2010). In detailed: Staining intensity was scored as follows: 1 (no staining), 2 (weak staining), 3 (moderate staining) and 4 (strong staining). The percentage of positively stained tumour cells was assessed as a proportion of the total number of tumour cells present in the section as follows: 1 (<20%), 2 (≥20–50%), 3 (>50–80%) and 4 (>80%). Then from staining intensity and percentage of positively stained cells, we created immunoreactivity score by multiplying staining intensity and the percentage of positivity. The score, therefore, ranged from 1 to 16 (divided into low (≤ 5), moderate (≥ 6 – 10) and high (≥11 – 16). However, specifications for staining intensity and the percentage of positivity used in this study are the standard system used by Pathology Anatomy Laboratory of Dr. Hasan Sadikin Hospital Bandung since 1990. These score systems are also adopted in some breast cancer diagnostic centres in Indonesia.

Immunoreactive score system used in this study have been published elsewhere (Thike et al., 2010). We mentioned this in Immunohistochemistry Section of our Methods.
In this study produce all raw data related to: type of TNBC (Basal like and Non Basal-like), subtype of cancer, expression of SOX2, P63 and CK 5/6 including staining intensity, distribution of the positively cells and immunoreactive score.

Reference:
Thike AA, Cheok PY, Jara-Lazaro AR, et al. Triple-negative breast cancer: clinicopathological characteristics and relationship with basal-like breast cancer. Mod Pathol. 2010;23(1):123–133.
Livasy CA, Karaca G, Nanda R, et al. Phenotypic evaluation of the basal-like subtype of invasive breast carcinoma. Mod Pathol 2006;19:264-271
Nielsen TO, Hsu FD, Jensen K, et al. Immunohistochemical and clinical characterization of the basal-like subtype of invasive breast carcinoma Clin Cancer Res, 2004;10:5367-5374
Rakha EA, El-Sayed ME, Green AR, et al. Breast carcinoma with basal differentiation: a proposal for pathology definition based on basal cytokeratin expression. Histopathology. 2007 Mar;50(4):434-8.
Sasa M, Bando Y, Takahashi M, et al. Screening for basal marker expression is necessary for decision of therapeutic strategy for triple-negative breast cancer. J Surg Oncol, 2008; 97: 30-34
Competing Interests: There is no competing of interest Close
Report a concern
Respond or Comment

COMMENTS ON THIS REPORT

Author Response 08 Jan 2018

Harapan Harapan, Medical Research Unit, School of Medicine, Syiah Kuala University, Banda Aceh, Indonesia

08 Jan 2018

Author Response

Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. We would like to ... Continue reading Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. We would like to confirm that predictive diagnostic analysis in this study was conducted using qualified statistician. There is no evidence to support the importance of the negative correlation between immunoexpression of P63 cytoplasm and immunoexpression SOX2 nucleus in metaplastic carcinoma, and therefore we are unable to discuss this finding in depth. In conclusion section, we have added some additional principal finding as suggested by the reviewer.
Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. We would like to confirm that predictive diagnostic analysis in this study was conducted using qualified statistician. There is no evidence to support the importance of the negative correlation between immunoexpression of P63 cytoplasm and immunoexpression SOX2 nucleus in metaplastic carcinoma, and therefore we are unable to discuss this finding in depth. In conclusion section, we have added some additional principal finding as suggested by the reviewer.
Competing Interests: There is no competing interest in this study. Close
Report a concern
Author Response 08 Jan 2018

Harapan Harapan, Medical Research Unit, School of Medicine, Syiah Kuala University, Banda Aceh, Indonesia

08 Jan 2018

Author Response

Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. In the revised manuscript, ... Continue reading Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. In the revised manuscript, the data of the incidence from global, Southeast Asian countries and Indonesia are included.

In our study, we used the histological paraffin blocks that have been confirmed as TNBC previously by testing ER, PR and HER2. To differentiate Basal like and non Basal like, either CK 5/6 or EGFR staining could be used with no significant different sensitivity and specificity (Livasy et al., 2006; Nielsen et al., 2004). In this study, CK 5/6 was employed to differentiate TNB into Basal like and non Basal like and this procedure was conducted during the study. This method is established method to define basal phenotype (Sasa et al., 2008; Rakha et al., 2007).

In our study, we used the histological paraffin blocks that have been confirmed as TNBC previously by testing ER, PR and HER2. To differentiate Basal like and non Basal like, either CK 5/6 or EGFR staining could be used with no significant different sensitivity and specificity (Livasy et al., 2006; Nielsen et al., 2004). In this study, CK 5/6 was employed to differentiate TNB into Basal like and non Basal like and this procedure was conducted during the study. This method is established method to define basal phenotype (Sasa et al., 2008; Rakha et al., 2007). CK 5/6 and EGFR.

As mentioned Immunohistochemistry Section, the principle of the scoring system for immunoexpression of P63 and SOX2 used in this study was have been elsewhere with modification (Thike et al., 2010). In detailed: Staining intensity was scored as follows: 1 (no staining), 2 (weak staining), 3 (moderate staining) and 4 (strong staining). The percentage of positively stained tumour cells was assessed as a proportion of the total number of tumour cells present in the section as follows: 1 (<20%), 2 (≥20–50%), 3 (>50–80%) and 4 (>80%). Then from staining intensity and percentage of positively stained cells, we created immunoreactivity score by multiplying staining intensity and the percentage of positivity. The score, therefore, ranged from 1 to 16 (divided into low (≤ 5), moderate (≥ 6 – 10) and high (≥11 – 16). However, specifications for staining intensity and the percentage of positivity used in this study are the standard system used by Pathology Anatomy Laboratory of Dr. Hasan Sadikin Hospital Bandung since 1990. These score systems are also adopted in some breast cancer diagnostic centres in Indonesia.

Immunoreactive score system used in this study have been published elsewhere (Thike et al., 2010). We mentioned this in Immunohistochemistry Section of our Methods.
In this study produce all raw data related to: type of TNBC (Basal like and Non Basal-like), subtype of cancer, expression of SOX2, P63 and CK 5/6 including staining intensity, distribution of the positively cells and immunoreactive score.

Reference:
Thike AA, Cheok PY, Jara-Lazaro AR, et al. Triple-negative breast cancer: clinicopathological characteristics and relationship with basal-like breast cancer. Mod Pathol. 2010;23(1):123–133.
Livasy CA, Karaca G, Nanda R, et al. Phenotypic evaluation of the basal-like subtype of invasive breast carcinoma. Mod Pathol 2006;19:264-271
Nielsen TO, Hsu FD, Jensen K, et al. Immunohistochemical and clinical characterization of the basal-like subtype of invasive breast carcinoma Clin Cancer Res, 2004;10:5367-5374
Rakha EA, El-Sayed ME, Green AR, et al. Breast carcinoma with basal differentiation: a proposal for pathology definition based on basal cytokeratin expression. Histopathology. 2007 Mar;50(4):434-8.
Sasa M, Bando Y, Takahashi M, et al. Screening for basal marker expression is necessary for decision of therapeutic strategy for triple-negative breast cancer. J Surg Oncol, 2008; 97: 30-34
Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. In the revised manuscript, the data of the incidence from global, Southeast Asian countries and Indonesia are included.

In our study, we used the histological paraffin blocks that have been confirmed as TNBC previously by testing ER, PR and HER2. To differentiate Basal like and non Basal like, either CK 5/6 or EGFR staining could be used with no significant different sensitivity and specificity (Livasy et al., 2006; Nielsen et al., 2004). In this study, CK 5/6 was employed to differentiate TNB into Basal like and non Basal like and this procedure was conducted during the study. This method is established method to define basal phenotype (Sasa et al., 2008; Rakha et al., 2007).

In our study, we used the histological paraffin blocks that have been confirmed as TNBC previously by testing ER, PR and HER2. To differentiate Basal like and non Basal like, either CK 5/6 or EGFR staining could be used with no significant different sensitivity and specificity (Livasy et al., 2006; Nielsen et al., 2004). In this study, CK 5/6 was employed to differentiate TNB into Basal like and non Basal like and this procedure was conducted during the study. This method is established method to define basal phenotype (Sasa et al., 2008; Rakha et al., 2007). CK 5/6 and EGFR.

As mentioned Immunohistochemistry Section, the principle of the scoring system for immunoexpression of P63 and SOX2 used in this study was have been elsewhere with modification (Thike et al., 2010). In detailed: Staining intensity was scored as follows: 1 (no staining), 2 (weak staining), 3 (moderate staining) and 4 (strong staining). The percentage of positively stained tumour cells was assessed as a proportion of the total number of tumour cells present in the section as follows: 1 (<20%), 2 (≥20–50%), 3 (>50–80%) and 4 (>80%). Then from staining intensity and percentage of positively stained cells, we created immunoreactivity score by multiplying staining intensity and the percentage of positivity. The score, therefore, ranged from 1 to 16 (divided into low (≤ 5), moderate (≥ 6 – 10) and high (≥11 – 16). However, specifications for staining intensity and the percentage of positivity used in this study are the standard system used by Pathology Anatomy Laboratory of Dr. Hasan Sadikin Hospital Bandung since 1990. These score systems are also adopted in some breast cancer diagnostic centres in Indonesia.

Immunoreactive score system used in this study have been published elsewhere (Thike et al., 2010). We mentioned this in Immunohistochemistry Section of our Methods.
In this study produce all raw data related to: type of TNBC (Basal like and Non Basal-like), subtype of cancer, expression of SOX2, P63 and CK 5/6 including staining intensity, distribution of the positively cells and immunoreactive score.

Reference:
Thike AA, Cheok PY, Jara-Lazaro AR, et al. Triple-negative breast cancer: clinicopathological characteristics and relationship with basal-like breast cancer. Mod Pathol. 2010;23(1):123–133.
Livasy CA, Karaca G, Nanda R, et al. Phenotypic evaluation of the basal-like subtype of invasive breast carcinoma. Mod Pathol 2006;19:264-271
Nielsen TO, Hsu FD, Jensen K, et al. Immunohistochemical and clinical characterization of the basal-like subtype of invasive breast carcinoma Clin Cancer Res, 2004;10:5367-5374
Rakha EA, El-Sayed ME, Green AR, et al. Breast carcinoma with basal differentiation: a proposal for pathology definition based on basal cytokeratin expression. Histopathology. 2007 Mar;50(4):434-8.
Sasa M, Bando Y, Takahashi M, et al. Screening for basal marker expression is necessary for decision of therapeutic strategy for triple-negative breast cancer. J Surg Oncol, 2008; 97: 30-34
Competing Interests: There is no competing of interest Close
Report a concern

Views

Reviewer Report 12 Oct 2017

Irianiwati Widodo, Department of Anatomical Pathology, Faculty of Medicine, Gadjah Mada University, Yogyakarta, Indonesia

Approved with Reservations

https://doi.org/10.5256/f1000research.13722.r26921

Introduction:

Do not explain the epidemiology of breast cancer too long, but explain more detail about TNBC and its subtypes (Basal-like and non basal-like), such as differences in histological type, behaviour, prognosis, treatment etc. So, the

Introduction:

Do not explain the epidemiology of breast cancer too long, but explain more detail about TNBC and its subtypes (Basal-like and non basal-like), such as differences in histological type, behaviour, prognosis, treatment etc. So, the role of p63 and SOX2 on TNBC becomes clearer.

Method:

Statistical analysis predictive diagnostic value of cytoplasmic p63 expression of basal-like and non basal-like TNBC, as far as I know there should be also negative p63 expression. Therefore I suggest to ask a qualified statistician.

Discussion:

Explain the importance of negative correlation between p63 expression and metaplastic carcinoma. Compare with previous studies.

Conclusion:

Mention also other important results of this study such as:

The frequency of non basal-like is higher than basal-like.
The most common histological type of TNBC is metaplastic carcinoma.
Negative correlation between p63 expression and metaplastic carcinoma
SOX2 expression in TNBC, even there is no stattistically significant

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Yes
Are sufficient details of methods and analysis provided to allow replication by others?

Yes
If applicable, is the statistical analysis and its interpretation appropriate?

I cannot comment. A qualified statistician is required.
Are all the source data underlying the results available to ensure full reproducibility?

Partly
Are the conclusions drawn adequately supported by the results?

Partly

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: I am a pathologist with major interest in breast cancer

CITE

Report a concern

Author Response 08 Jan 2018

Harapan Harapan, Medical Research Unit, School of Medicine, Syiah Kuala University, Banda Aceh, Indonesia

08 Jan 2018

Author Response

Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. We would like to ... Continue reading Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. We would like to confirm that predictive diagnostic analysis in this study was conducted using qualified statistician. There is no evidence to support the importance of the negative correlation between immunoexpression of P63 cytoplasm and immunoexpression SOX2 nucleus in metaplastic carcinoma, and therefore we are unable to discuss this finding in depth. In conclusion section, we have added some additional principal finding as suggested by the reviewer.
Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. We would like to confirm that predictive diagnostic analysis in this study was conducted using qualified statistician. There is no evidence to support the importance of the negative correlation between immunoexpression of P63 cytoplasm and immunoexpression SOX2 nucleus in metaplastic carcinoma, and therefore we are unable to discuss this finding in depth. In conclusion section, we have added some additional principal finding as suggested by the reviewer.
Competing Interests: No competing interests were disclosed. Close
Report a concern
Respond or Comment

COMMENTS ON THIS REPORT

Author Response 08 Jan 2018

Harapan Harapan, Medical Research Unit, School of Medicine, Syiah Kuala University, Banda Aceh, Indonesia

08 Jan 2018

Author Response

Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. We would like to ... Continue reading Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. We would like to confirm that predictive diagnostic analysis in this study was conducted using qualified statistician. There is no evidence to support the importance of the negative correlation between immunoexpression of P63 cytoplasm and immunoexpression SOX2 nucleus in metaplastic carcinoma, and therefore we are unable to discuss this finding in depth. In conclusion section, we have added some additional principal finding as suggested by the reviewer.
Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. We would like to confirm that predictive diagnostic analysis in this study was conducted using qualified statistician. There is no evidence to support the importance of the negative correlation between immunoexpression of P63 cytoplasm and immunoexpression SOX2 nucleus in metaplastic carcinoma, and therefore we are unable to discuss this finding in depth. In conclusion section, we have added some additional principal finding as suggested by the reviewer.
Competing Interests: No competing interests were disclosed. Close
Report a concern

Comments on this article Comments (0)

Version 2

VERSION 2 PUBLISHED 28 Sep 2017

Open Peer Review

Reviewer Status

Reviewer Reports

	Invited Reviewers
	1	2
Version 2 (revision) 08 Jan 18	read	read
Version 1 28 Sep 17	read	read

Irianiwati Widodo, Gadjah Mada University, Yogyakarta, Indonesia
Diah Rini Handjari, University of Indonesia, Jakarta, Indonesia

Comments on this article

All Comments(0)

Add a comment

Browse by related subjects

Back to all reports

Reviewer Report

4 Views

20 Feb 2018 | for Version 2

Diah Rini Handjari, University of Indonesia, Jakarta, Indonesia

4 Views Cite this report Responses(0)

Approved

This revised version is good and sufficient for indexing.

Competing Interests

No competing interests were disclosed.

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.

Respond to this report

Responses (0)

Back to all reports

Reviewer Report

11 Views

06 Feb 2018 | for Version 2

Irianiwati Widodo, Department of Anatomical Pathology, Faculty of Medicine, Gadjah Mada University, Yogyakarta, Indonesia

11 Views Cite this report Responses(0)

Approved

This revised version is good and already sufficient for publication.

Competing Interests

No competing interests were disclosed.

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.

Respond to this report

Responses (0)

Back to all reports

Reviewer Report

14 Views

20 Oct 2017 | for Version 1

Diah Rini Handjari, University of Indonesia, Jakarta, Indonesia

14 Views Cite this report Responses(2)

Approved With Reservations

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Yes
Are sufficient details of methods and analysis provided to allow replication by others?

Yes
If applicable, is the statistical analysis and its interpretation appropriate?

I cannot comment. A qualified statistician is required.
Are all the source data underlying the results available to ensure full reproducibility?

Partly
Are the conclusions drawn adequately supported by the results?

Partly

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Pathologist with major interest in colorectal cancer

Respond to this report

Responses (2)

Author Response

08 Jan 2018

Harapan Harapan, Medical Research Unit, School of Medicine, Syiah Kuala University, Banda Aceh, Indonesia

Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. We would like to confirm that predictive diagnostic analysis in this study was conducted using qualified statistician. There is no evidence to support the importance of the negative correlation between immunoexpression of P63 cytoplasm and immunoexpression SOX2 nucleus in metaplastic carcinoma, and therefore we are unable to discuss this finding in depth. In conclusion section, we have added some additional principal finding as suggested by the reviewer.

View more View less

Competing Interests

There is no competing interest in this study.

Author Response

08 Jan 2018

Harapan Harapan, Medical Research Unit, School of Medicine, Syiah Kuala University, Banda Aceh, Indonesia

Thank you for comments and suggestion. We have deleted some sentences related to the general introduction of breast cancer and added specific information related to TNBC. In the revised manuscript, the data of the incidence from global, Southeast Asian countries and Indonesia are included.

In our study, we used the histological paraffin blocks that have been confirmed as TNBC previously by testing ER, PR and HER2. To differentiate Basal like and non Basal like, either CK 5/6 or EGFR staining could be used with no significant different sensitivity and specificity (Livasy et al., 2006; Nielsen et al., 2004). In this study, CK 5/6 was employed to differentiate TNB into Basal like and non Basal like and this procedure was conducted during the study. This method is established method to define basal phenotype (Sasa et al., 2008; Rakha et al., 2007).

In our study, we used the histological paraffin blocks that have been confirmed as TNBC previously by testing ER, PR and HER2. To differentiate Basal like and non Basal like, either CK 5/6 or EGFR staining could be used with no significant different sensitivity and specificity (Livasy et al., 2006; Nielsen et al., 2004). In this study, CK 5/6 was employed to differentiate TNB into Basal like and non Basal like and this procedure was conducted during the study. This method is established method to define basal phenotype (Sasa et al., 2008; Rakha et al., 2007). CK 5/6 and EGFR.

As mentioned Immunohistochemistry Section, the principle of the scoring system for immunoexpression of P63 and SOX2 used in this study was have been elsewhere with modification (Thike et al., 2010). In detailed: Staining intensity was scored as follows: 1 (no staining), 2 (weak staining), 3 (moderate staining) and 4 (strong staining). The percentage of positively stained tumour cells was assessed as a proportion of the total number of tumour cells present in the section as follows: 1 (<20%), 2 (≥20–50%), 3 (>50–80%) and 4 (>80%). Then from staining intensity and percentage of positively stained cells, we created immunoreactivity score by multiplying staining intensity and the percentage of positivity. The score, therefore, ranged from 1 to 16 (divided into low (≤ 5), moderate (≥ 6 – 10) and high (≥11 – 16). However, specifications for staining intensity and the percentage of positivity used in this study are the standard system used by Pathology Anatomy Laboratory of Dr. Hasan Sadikin Hospital Bandung since 1990. These score systems are also adopted in some breast cancer diagnostic centres in Indonesia.

Immunoreactive score system used in this study have been published elsewhere (Thike et al., 2010). We mentioned this in Immunohistochemistry Section of our Methods.
In this study produce all raw data related to: type of TNBC (Basal like and Non Basal-like), subtype of cancer, expression of SOX2, P63 and CK 5/6 including staining intensity, distribution of the positively cells and immunoreactive score.

Reference:
Thike AA, Cheok PY, Jara-Lazaro AR, et al. Triple-negative breast cancer: clinicopathological characteristics and relationship with basal-like breast cancer. Mod Pathol. 2010;23(1):123–133.
Livasy CA, Karaca G, Nanda R, et al. Phenotypic evaluation of the basal-like subtype of invasive breast carcinoma. Mod Pathol 2006;19:264-271
Nielsen TO, Hsu FD, Jensen K, et al. Immunohistochemical and clinical characterization of the basal-like subtype of invasive breast carcinoma Clin Cancer Res, 2004;10:5367-5374
Rakha EA, El-Sayed ME, Green AR, et al. Breast carcinoma with basal differentiation: a proposal for pathology definition based on basal cytokeratin expression. Histopathology. 2007 Mar;50(4):434-8.
Sasa M, Bando Y, Takahashi M, et al. Screening for basal marker expression is necessary for decision of therapeutic strategy for triple-negative breast cancer. J Surg Oncol, 2008; 97: 30-34

View more View less

Competing Interests

There is no competing of interest

Back to all reports

Reviewer Report

18 Views

12 Oct 2017 | for Version 1

Irianiwati Widodo, Department of Anatomical Pathology, Faculty of Medicine, Gadjah Mada University, Yogyakarta, Indonesia

18 Views Cite this report Responses(1)

Approved With Reservations

Introduction:

Do not explain the epidemiology of breast cancer too long, but explain more detail about TNBC and its subtypes (Basal-like and non basal-like), such as differences in histological type, behaviour, prognosis, treatment etc. So, the role of p63 and SOX2 on TNBC becomes clearer.

Method:

Statistical analysis predictive diagnostic value of cytoplasmic p63 expression of basal-like and non basal-like TNBC, as far as I know there should be also negative p63 expression. Therefore I suggest to ask a qualified statistician.

Discussion:

Explain the importance of negative correlation between p63 expression and metaplastic carcinoma. Compare with previous studies.

Conclusion:

Mention also other important results of this study such as:

The frequency of non basal-like is higher than basal-like.
The most common histological type of TNBC is metaplastic carcinoma.
Negative correlation between p63 expression and metaplastic carcinoma
SOX2 expression in TNBC, even there is no stattistically significant

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Yes
Are sufficient details of methods and analysis provided to allow replication by others?

Yes
If applicable, is the statistical analysis and its interpretation appropriate?

I cannot comment. A qualified statistician is required.
Are all the source data underlying the results available to ensure full reproducibility?

Partly
Are the conclusions drawn adequately supported by the results?

Partly

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

I am a pathologist with major interest in breast cancer

Respond to this report

Responses (1)

Author Response

08 Jan 2018

Harapan Harapan, Medical Research Unit, School of Medicine, Syiah Kuala University, Banda Aceh, Indonesia

View more View less

Competing Interests

No competing interests were disclosed.

Alongside their report, reviewers assign a status to the article:

Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested

Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit.

Not approved - fundamental flaws in the paper seriously undermine the findings and conclusions

Click here to access the data.

Downloaded data do not display as expected? Download the data

[1] 1. Key TJ, Verkasalo PK, Banks E: Epidemiology of breast cancer. Lancet Oncol. 2001; 2(3): 133–140. PubMed Abstract | Publisher Full Text

[2] 2. Torre LA, Bray F, Siegel RL, et al.: Global cancer statistics, 2012. CA Cancer J Clin. 2015; 65(2): 87–108. PubMed Abstract | Publisher Full Text

[3] 3. Althuis MD, Dozier JM, Anderson WF, et al.: Global trends in breast cancer incidence and mortality 1973–1997. Int J Epidemiol. 2005; 34(2): 405–412. PubMed Abstract | Publisher Full Text

[4] 4. Ghoncheh M, Momenimovahed Z, Salehiniya H: Epidemiology, Incidence and Mortality of Breast Cancer in Asia. Asian Pac J Cancer Prev. 2016; 17(S3): 47–52. PubMed Abstract | Publisher Full Text

[5] 5. Plasilova ML, Hayse B, Killelea BK, et al.: Features of triple-negative breast cancer: Analysis of 38,813 cases from the national cancer database. Medicine (Baltimore). 2016; 95(35): e4614. PubMed Abstract | Publisher Full Text | Free Full Text

[6] 6. Alcantara VS, Lim GH, Lim SH, et al.: Incidence and prognosis of non-metastatic triple negative breast cancer (TNBC) among different races in Southeast Asia. J Surg Oncol. 2017; 115(5): 523–537. PubMed Abstract | Publisher Full Text

[7] 7. Kusumadjayanti N, Badudu DF, Hernowo BS: Characteristics of patients with estrogen receptor (ER)-negative, progesterone receptor (PR)-negative, and HER2-negative invasive breast cancer in Dr. Hasan Sadikin General Hospital, Bandung, Indonesia from 2010 to 2011. Althea Med J. 2015; 2(3): 391–394. Publisher Full Text

[8] 8. Cleator S, Heller W, Coombes RC: Triple-negative breast cancer: therapeutic options. Lancet Oncol. 2007; 8(3): 235–44. PubMed Abstract | Publisher Full Text

[9] 9. Kobayashi S: Basal-like subtype of breast cancer: a review of its unique characteristics and their clinical significance. Breast Cancer. 2008; 15(2): 153–8. PubMed Abstract | Publisher Full Text

[10] 10. Rakha EA, Reis-Filho JS, Ellis IO: Basal-like breast cancer: a critical review. J Clin Oncol. 2008; 26(15): 2568–2581. PubMed Abstract | Publisher Full Text

[11] 11. AIHW: Breast cancer survival by size and nodal status in Australia. In: Registries NBCCAAoC, ed. Cancer series no. 39. Canberra: AIHW; 2007. Reference Source

[12] 12. Allemani C, Minicozzi P, Berrino F, et al.: Predictions of survival up to 10 years after diagnosis for European women with breast cancer in 2000–2002. Int J Cancer. 2013; 132(10): 2404–2412. PubMed Abstract | Publisher Full Text

[13] 13. Allemani C, Weir HK, Carreira H, et al.: Global surveillance of cancer survival 1995–2009: analysis of individual data for 25,676,887 patients from 279 population-based registries in 67 countries (CONCORD-2). Lancet. 2015; 385(9972): 977–1010. PubMed Abstract | Publisher Full Text | Free Full Text

[14] 14. Naroda S, Iqbala J, Miller AB: Why have breast cancer mortality rates declined? J Cancer Policy. 2015; 5: 8–17. Publisher Full Text

[15] 15. Reisenbichler ES, Ross JR, Hameed O: The clinical use of a P63/cytokeratin7/18/cytokeratin5/14 antibody cocktail in diagnostic breast pathology. Ann Diagn Pathol. 2014; 18(6): 313–318. PubMed Abstract | Publisher Full Text

[16] 16. Liu Y, Coates PJ, Nenutil R, et al.: Lack of correlation between markers of breast cancer initiating cells. Breast Cancer Res. 2010; 12(Suppl 1): P38. Publisher Full Text | Free Full Text

[17] 17. Koker MM, Kleer CG: p63 expression in breast cancer: a highly sensitive and specific marker of metaplastic carcinoma. Am J Surg Pathol. 2004; 28(11): 1506–1512. PubMed Abstract | Publisher Full Text

[18] 18. Assi HA, Khoury KE, Dbouk H, et al.: Epidemiology and prognosis of breast cancer in young women. J Thorac Dis. 2013; 5 Suppl 1: S2–8. PubMed Abstract | Publisher Full Text | Free Full Text

[19] 19. Flores ER, Sengupta S, Miller JB, et al.: Tumor predisposition in mice mutant for p63 and p73: evidence for broader tumor suppressor functions for the p53 family. Cancer Cell. 2005; 7(4): 363–373. PubMed Abstract | Publisher Full Text

[20] 20. Su X, Chakravarti D, Cho MS, et al.: TAp63 suppresses metastasis through coordinate regulation of Dicer and miRNAs. Nature. 2010; 467(7318): 986–990. PubMed Abstract | Publisher Full Text | Free Full Text

[21] 21. Bergholz J, Xiao ZX: Role of p63 in Development, Tumorigenesis and Cancer Progression. Cancer Microenviron. 2012; 5(3): 311–322. PubMed Abstract | Publisher Full Text | Free Full Text

[22] 22. Thike AA, Cheok PY, Jara-Lazaro AR, et al.: Triple-negative breast cancer: clinicopathological characteristics and relationship with basal-like breast cancer. Mod Pathol. 2010; 23(1): 123–133. PubMed Abstract | Publisher Full Text

[23] 23. Harton AM, Wang HH, Schnitt SJ, et al.: p63 Immunocytochemistry improves accuracy of diagnosis with fine-needle aspiration of the breast. Am J Clin Pathol. 2007; 128(1): 80–85. PubMed Abstract | Publisher Full Text

[24] 24. Weina K, Utikal J: SOX2 and cancer: current research and its implications in the clinic. Clin Transl Med. 2014; 3: 19. PubMed Abstract | Publisher Full Text | Free Full Text

[25] 25. Sarkar A, Hochedlinger K: The sox family of transcription factors: versatile regulators of stem and progenitor cell fate. Cell Stem Cell. 2013; 12(1): 15–30. PubMed Abstract | Publisher Full Text | Free Full Text

[26] 26. Stolzenburg S, Rots MG, Beltran AS, et al.: Targeted silencing of the oncogenic transcription factor SOX2 in breast cancer. Nucleic Acids Res. 2012; 40(14): 6725–6740. PubMed Abstract | Publisher Full Text | Free Full Text

[27] 27. Leis O, Eguiara A, Lopez-Arribillaga E, et al.: Sox2 expression in breast tumours and activation in breast cancer stem cells. Oncogene. 2012; 31(11): 1354–1365. PubMed Abstract | Publisher Full Text

[28] 28. Piva M, Domenici G, Iriondo O, et al.: Sox2 promotes tamoxifen resistance in breast cancer cells. EMBO Mol Med. 2014; 6(1): 66–79. PubMed Abstract | Publisher Full Text | Free Full Text

[29] 29. Anderson G, Bancroft J: Tissue processing and microtomy. In: Bancroft JG, M., ed. Theory and practiceal of histological techniques 5th Edition. Edinburgh Churchill Livingstone; 2002; 109–123.

[30] 30. Matkovic B, Juretic A, Separovic V, et al.: Immunohistochemical analysis of ER, PR, HER-2, CK 5/6, p63 and EGFR antigen expression in medullary breast cancer. Tumori. 2008; 94(6): 838–844. PubMed Abstract

[31] 31. Tacha DE, Bloom K, Kyshtoobayava A, et al.: A double immunostaining technique with a cocktail CK5, CK14, p63, CK7 and CK18 distinguishes between hyperplasia of the usual type, atypical hyperplasia, microinvasive and basal phenotype breast cancers. Modern Pathology. 2009; 22: 388a.

[32] 32. Liu K, Xie F, Gao A, et al.: SOX2 regulates multiple malignant processes of breast cancer development through the SOX2/miR-181a-5p, miR-30e-5p/TUSC3 axis. Mol Cancer. 2017; 16(1): 62. PubMed Abstract | Publisher Full Text | Free Full Text

[33] 33. Lengerke C, Fehm T, Kurth R, et al.: Expression of the embryonic stem cell marker SOX2 in early-stage breast carcinoma. BMC Cancer. 2011; 11: 42. PubMed Abstract | Publisher Full Text | Free Full Text

[34] 34. Sun Y, Zhang R, Wang MJ, et al.: SOX2 autoantibodies as noninvasive serum biomarker for breast carcinoma. Cancer Epidemiol Biomarkers Prev. 2012; 21(11): 2043–2047. PubMed Abstract | Publisher Full Text

[35] 35. Zheng Y, Qin B, Li F, et al.: Clinicopathological significance of Sox2 expression in patients with breast cancer: a meta-analysis. Int J Clin Exp Med. 2015; 8(12): 22382–22392. PubMed Abstract | Free Full Text

[36] 36. Reno KK, Muhammad NDL, et al.: Dataset 1 in: Immunoexpression of P63 and SOX2 in triple-negative breast cancers, Indonesia. F1000Research. 2017. Data Source

Immunoexpression of P63 and SOX2 in triple-negative breast cancers, Indonesia

Abstract

Keywords

Revised Amendments from Version 1

Introduction

Methods

Study setting and histological slides

Immunohistochemistry

Statistical analysis

Results

Clinicopathology and classification of TNBC

Table 1. Histopathology types of TNBC samples used in this study (N=40).

Immunoreactivity score of P63 and SOX2

Table 2. Immunoreactivity score of P63 and SOX2 in TNBC.

Correlation between immunoexpression of P63 and SOX2

Table 3. Correlation between immunoexpression of P63 cytoplasm and SOX2 in Basal-like type TNBC.

Immunoexpression of P63 and SOX2 in Basal-like and Non basal-like type of TNBC

Table 4. Immunoexpression of P63 and SOX2 in Basal-like type TNBC and Non basal-like type TNBC.

Predictor diagnostic value of P63 in the cytoplasm for diagnosing Basal like type TNBC

Table 5. Predictor diagnostic value of P63 in the cytoplasm for diagnosing Basal-like type TNBC.

Figure 1. Receiver operating curve of P63 cytoplasm immunoexpression for determining TNBC basal-like type.

Discussion

Conclusions

Data availability

Competing interests

Grant information

Acknowledgments

References

Comments on this article Comments (0)

Open Peer Review

Comments on this article Comments (0)

Open Peer Review

Reviewer Status

Reviewer Reports

Comments on this article

Browse by related subjects

The problem

How to fix it

Competing Interests Policy

Stay Updated