Protocol for the evaluation of cyclin D1 expression in tumor tissue and surgical margins in oral squamous cell carcinoma – a retrospective study

ANKITA CHAVHAN; Swati Patil; Madhuri Gawande; Alka Hande; Archana Sonone; Aayushi Pakhale

doi:10.12688/f1000research.134525.1

Home Browse Protocol for the evaluation of cyclin D1 expression in tumor tissue...

ALL Metrics

-

Views

-

Downloads

Get PDF

Get XML

Export

▬

✚

Study Protocol

Protocol for the evaluation of cyclin D1 expression in tumor tissue and surgical margins in oral squamous cell carcinoma – a retrospective study

[version 1; peer review: 1 not approved]

ANKITA CHAVHAN ¹, Swati Patil², Madhuri Gawande², Alka Hande ², Archana Sonone ², Aayushi Pakhale ²

ANKITA CHAVHAN ¹, Swati Patil², [...] Madhuri Gawande², Alka Hande ², Archana Sonone ², Aayushi Pakhale ²

PUBLISHED 01 Sep 2023

Author details Author details

¹ “Department of Oral & Maxillofacial Pathology and Microbiology ”, Post graduate student, Sharad Pawar Dental College & Hospital, Datta Meghe Institute of Higher Education and Research, “Sawangi (Meghe)”, Wardha, Wardha, Maharashtra, 442004, India
² “Department of Oral & Maxillofacial Pathology and Microbiology”, “Sharad Pawar Dental College & Hospital”, Professor, “Datta Meghe Institute of Higher Education and Research , Sawangi (Meghe), Wardha, Wardha, Maharashtra, 442004, India

ANKITA CHAVHAN
Roles: Conceptualization, Formal Analysis, Funding Acquisition, Investigation, Methodology, Project Administration, Resources, Software, Supervision, Validation, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing

Swati Patil
Roles: Supervision, Validation, Visualization

Madhuri Gawande
Roles: Supervision, Validation, Visualization

Alka Hande
Roles: Supervision, Validation, Visualization

Archana Sonone
Roles: Supervision, Validation, Visualization

Aayushi Pakhale
Roles: Supervision, Validation, Visualization

OPEN PEER REVIEW

REVIEWER STATUS

This article is included in the Datta Meghe Institute of Higher Education and Research collection.

Abstract

Oral squamous cell carcinoma (OSCC) is a malignant epithelial tumor with poor prognosis. There is an increase in the proliferation of epithelial cells in OSCC which is control by cell cycle regulators like Cyclin D1. Cyclin D1 is linked to a poor prognosis for patients, including aggressive malignancies, recurrence, protracted remission, etc.
To evaluate Cyclin D1 immunoexpression in different grades and surgical margins in OSCC. All tissue sections will be processed for Cyclin D1 IHC. After that light microscopic evaluation of Cyclin D1 expression was classified as weak (scoring 1-4), moderate (score 5-8), or strong based on a final expression score that was obtained by multiplying the labelling index score by the intensity score (score 9–12).
The present study will find the immunohistochemical expression of Cyclin D1 in OSCC. There will be variation in the immunoexpression of Cyclin D1 among different grades of OSCC its surgical margin and normal oral mucosa.
We hypothesize that: As the disease condition goes towards poor grade, intensity of cyclin D1 expression immunohistochemically may increase. Hence, the intensity can be used as a prognostic marker.

Keywords

Cyclin d1, Cell proliferation,Oral potential malignant disorder, Oral squamous cell carcinoma, Immunohistochemistry

Corresponding authors: ANKITA CHAVHAN, Swati Patil, Madhuri Gawande, Alka Hande, Archana Sonone, Aayushi Pakhale

Competing interests: No competing interests were disclosed.

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

Copyright: © 2023 CHAVHAN A 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.

How to cite: CHAVHAN A, Patil S, Gawande M et al. Protocol for the evaluation of cyclin D1 expression in tumor tissue and surgical margins in oral squamous cell carcinoma – a retrospective study [version 1; peer review: 1 not approved]. F1000Research 2023, 12:1096 (https://doi.org/10.12688/f1000research.134525.1) First published: 01 Sep 2023, 12:1096 (https://doi.org/10.12688/f1000research.134525.1) Latest published: 01 Sep 2023, 12:1096 (https://doi.org/10.12688/f1000research.134525.1)

Introduction

Background and rationale

Oral squamous cell carcinoma (OSCC) is a malignant epithelial tumor with poor prognosis. OSCC is one of the top causes of death in India and Southeast Asia.¹ Oral squamous cell carcinoma caused due to chewing tobacco, such as panmasala, gutkha, gudakhu, khaini, and other products composed of tobacco.²^,³ Patients with middle-aged to older ages typically affected by OSCC.⁴ OSCC have a high rate of proliferation of malignant epithelial cells. The prognosis of OSCC depends on recommended surgical margin.

In the surgical margin, the epithelial cells may be altered by field cancerization. There is an increase in the proliferation of epithelial cells in OSCC which is control by cell cycle regulators like Cyclin D1.⁵

Cyclin control the orderly phases of the cells through the G1, S, G2, and M phases of the cell cycle.⁶^,⁷ It plays a role in cell cycle induction, transcriptional control, and DNA damage-induced death.² (CCND1) Cyclin D1, 11q13 location of chromosomes, carries the cell cycle from the G1 to the S phase by encoding a vital cell cycle regulating protein.⁸^‐¹⁰ The control of the cell cycle is facilitated by Cyclin D1, proto-oncogenes and an enzyme known as Cylin-dependent kinase 4 (CDK4). It also interacts with other transcriptional factor (TFs) and controls the activity of transcriptional cofactors.⁴ Lack of regulation of the cell cycle mechanism is an essential phase in the carcinogenic change. Cyclin D1 is linked to a poor prognosis for patients, including aggressive malignancies, recurrence, protracted remission, etc.³

Objectives

1) To evaluate the Cyclin D1 immuno-expression in –
- • Normal oral mucosa.
- • Different grades of OSCC.
- • Surgical margin of OSCC.
2) To compare Cyclin D1 immuno-expressions in –
- • Normal oral mucosa.
- • Different grades of OSCC.
- • Surgical margin of OSCC.

TRIAL DESIGN: Retrospective study.

Methods

Study Setting

This study will be conducted in the Department of Oral Pathology and Microbiology, Sharad Pawar Dental College in collaboration with Central Research Laboratory, Datta Meghe Institute of Higher Education & Reasearch, Sawangi (Meghe) Wardha.

Eligibility Criteria

Inclusion criteria

• Paraffin embedded tissue sections of well differentiated squamous cell carcinoma (WDSCC), moderately differentiated squamous cell carcinoma (MDSCC), poorly differentiated squamous cell carcinoma (PDSCC) histopathologically, their surgical margins and normal oral mucosa (NOM) will be retrieved from archival of department.
• 80 surgically treated patients who have been clinically and histopathologically diagnosed with OSCC.
• Recorded demographic data of clinical presentation, habits and its duration, histopathological findings, operative details will be noted in detail from department.
• Follow-up data for 2-3 years of disease-free survival.

Exclusion criteria

• Patients who have a history of pre-malignant lesion, recurrent or distant metastasis and pre-operative chemotherapy, radiation therapy and the patients previously having undergone surgery for recurrence (except biopsy).

Intervention

All tissue sections will be proceeded for Cyclin D1 IHC. After that, a light microscopic evaluation of Cyclin D1 expression was classified as weak (scoring 1-4), moderate (score 5-8), or strong based on a final expression score that was obtained by multiplying the labelling an index score by the intensity score (score 9–12).

Outcomes

Primary outcome

The present study will find the immunohistochemical expression of cyclin D1 tumor tissue and surgical margins of OSCC, in order to evaluate the prognosis of OSCC.

Secondary outcome

There will be a variation in the immunoexpression of Cyclin D1 among different grades of oral squamous cell carcinoma, its surgical margin and normal oral mucosa.

Sample Size

Considering the prevalence of OSCC as 70.7% in the outpatient department of Oral Pathology and Microbiology, using a single proportion formula, the sample size is calculated via the expression below.¹¹

n \geq Z {^{2}}_{1 - α / 2}^{*} p^{*} (1 - p) / d^{2}

Where,

Z²_{1-α/2 -} The level of significance at 5 %

i.e. 95 % confidence interval =1.96

p - Sample demonstrating +ve Cycline D1 expressions in tiny groups of cells in the basal layer of the

epithelium = 70.7% = 0.707

E - Errors of margins = 10 % = 0.10

n = 1.96²x 0.707 x (1-0.707) /0.10²

n = 80

Formula reference - Angadi PV, Krishnapillai R. Cyclin D1 expression in OSCC and verrucous carcinoma: correlation with histological differentiation. OS, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology. 2007 Mar 1;103(3):e30-5

Immunostaining

A paraffin block having suitable mass of tumor and acceptable amount of normal tissue is selected. On Poly-L-Lysinecoated slides, sections of having thickness of three micrometers are cut & placed. For De-paraffinization, sections are placed in the xylene solution. Sections are rehydrated by subjecting them to descending concentrations of alcohol (70-100%). In order to wash sections, tap water is used. Washing time for sections in distilled water is 60 seconds. After washing all the sections are transferred to Coplin’s jar containing the retrieval buffer solution. The solution which is used for Antigen retrieval is composed of 30 ml of retrieval solution in 1500 mili-liters of distilled water for fifteen to twenty minutes in the pressure cooker. Cooling is done at room temperature.

Sections are dipped once in distilled water. Sections are washed with Tris buffer solution for at least 5 minutes at room temperature. This step is repeated thrice. For peroxidase blocking a mixture of “3,5 Hydrogen Peroxide and Methanol” is used for thirty minutes. Tris buffer solution is used for washing the sections 3 times for 5 minutes each. Primary antibody cyclin D1 is applied at room temperature for 1 hour. Envision technique is performed by utilizing a labelled polymer for 30 minutes at room temperature. For washing the sections tris buffer solution is used thrice for five minutes.

The application of “DAB (3,3’-diaminobenzidine)” substrate is done for fifty to twenty minutes. The working DAB solution is comprised of following –one millilitre of DAB buffer & twenty five microliters of “DAB concentrate”. This time, washing of sections is done by Tris buffer, for 15-20 minutes. Sections are cleaned in distilled water. For counter staining Mayer’s hematoxylin is used, which is done for five minutes. Again, the washing of sections is done under tap water and then dried. Once the sections are dried; mounte in DPX. After that a microscopic examination is done. Calculations will be made to determine the median percentage per slide (labelling index) and the proportion of IHC positive tumour cells per hot spot. Labelling indices of 1-35%, 36-70%, and 71-100%, respectively, each received a score of 1, 2, or 3. According to their microscopic appearance, Cyclin D1 immunostaining intensity will be assigning each a score of 0, 1, 2, or 3 will indicate whether it is weak, intermediate, or strong. Cyclin D1 expression was classified as weak (scoring 1-4), moderate (score 5-8), or strong based on a final expression score that was obtained by multiplying the labelling index score by the intensity score (score 9–12). We will tabulate and do the necessary statistical analysis on all relevant clinical, histopathological, and immunohistochemical information that we will gather.

Dissemination

Will be published in an indexed journal.

Study status

Not started yet.

Discussion

King Y. Lam at el., (2000) examined how Cyclin D1 relates to the clinical and pathological characteristics of OSCC. They found that Cyclin D1 expressions were present in 63% of OSCC; that was typically modest but more commonly positive in high-grade lesions. Radiation therapy had no effect on either Cyclin D1's expression. Expression was unrelated to patient age, gender, survival, tumour stage, or patient demographics. Patients with the floor of the mouth, gingiva, oropharynx, and palate squamous cell carcinomas had overexpressed Cyclin D1. The researchers came to the conclusion that Cyclin D1 was often expressed in OSCC, connected to the high grade of tumours, and was not similar in different regions of the oral cavity, indicating diverse tumour biology of carcinoma from theis area.

R Miyamoto et al., in 2003 showed that the relationship among Cyclin D1 (CCND1) extension and over expressions in OSCCs were examined, and the predictive validity of Fluorescence in-situ Hybridization (FISH) on Cyclin D1 (CCND1) amplification with immunohistochemical over expression in fine-needle aspiration (FNA) samples was established. In every instance where CCND1 was amplified, they noticed CCND1 overexpression. While 14 of the 28 instances 50.0% that did not show such intensification showed CCND1 overexpression. They noticed insignificant The statistical correspondence among CCND1 over-expression and reduced endurance as well as the independent prognostic significance of CCND1 amplification and nodal status. They came to the conclusion that there is a need to look into the additional routes besides CCND1 amplification that control CCND1 expression. They also came to the conclusion that in OSCCs, CCND1 intensification is a more accurate analytical predictor than CCND1 over expression.

S Saawarn et al., in Cyclin D1's immunohistochemistry reactivity and expression, as well as their relationships to the location, clinically staged, and histological discrimination of OSCC, were examined in 2011. (OSCC). 45% of people showed expression of cyclin D1 of Oral SCC cases, although it was unrelated to the site or clinical stage. Moreover, they find a statistical significance link among the highest expression being detected in well-differentiated squamous cell carcinomas, followed by MDSCC and PDSCC. They came to the conclusion that as differentiation rises, Cyclin D1 expression does as well.

Y OHNISHI et al., in 2014 immunohistochemically examined the relationship among cyclin D1 expression and ki-67 expressions in oral Squamous CC and connected their expression among cells segregation, cells abundance, and metastasis. They discovered the expression of Cyclin D1 and Ki-67 in the nucleus of all 35 Squamous CC tissue, although they did not discover a connection between oral SCC differentiation and Cyclin D1 protein expression. Also, they discovered that 90% of metastatic foci had significant Cyclin D1 expression, in contrast to metastatic foci treated with preoperative adjuvant therapy, which had poor expression. Moreover, Cyclin D1 and Ki-67 were found in the basal to suprabasal cell layer of the WD oral SCC, while the highly-differentiated region also contained cyclin D1-positive and Ki-67-negative cells. Also, they proposed that in WD oral SCC, Cyclin D1 protein expression is essential for the growth and differentiation of cells.

Ethical considerations

Ethical approval received from Datta Meghe Institute of Higher Education and Research, Sawangi, Wardha.

IEC reference number- DMIHER (DU)/IEC/2023/842.

Informed written consent was obtained from the patient before surgery and before the before the write-up of the protocol as per the data requirement of this study.

Data availability

No data associated with this study.

Reporting guidelines

Zenodo, STROBE checklist for cross sectional study.

DOI - 10.5281/zenodo.7874225.

Data are available under the terms of the Creative Commons Attribution 4.0 International license (CC-BY 4.0).

Acknowledgements

I would like to thank my institute and my colleagues.

References

1. Sherkheli MA, Gisselmann G, Hatt H: Supercooling agent icilin blocks a warmth-sensing ion channel TRPV3. Sci. World J. 2012 Jan 1; 2012: 1–8. PubMed Abstract | Publisher Full Text | Free Full Text
2. Ohnishi Y, Watanabe M, Wato M, et al.: Cyclin D1 expression is correlated with cell differentiation and cell proliferation in oral squamous cell carcinomas. Oncol. Lett. 2014 Apr 1; 7(4): 1123–1127. PubMed Abstract | Publisher Full Text | Free Full Text
3. Mishra R, Das BR: Cyclin D1 expression and its possible regulation in chewing tobacco mediated oral squamous cell carcinoma progression. Arch. Oral Biol. 2009 Oct 1; 54(10): 917–923. PubMed Abstract | Publisher Full Text
4. Moharil RB, Khandekar S, Dive A, et al.: Cyclin D1 in oral premalignant lesions and oral squamous cell carcinoma: An immunohistochemical study. Journal of Oral and Maxillofacial Pathology: JOMFP. 2020 May; 24(2): 397. PubMed Abstract | Publisher Full Text | Free Full Text
5. Lam KY, Ng IO, Yuen AP, et al.: Cyclin D1 expression in oral squamous cell carcinomas: clinicopathological relevance and correlation with p53 expression. J. Oral Pathol. Med. 2000 Apr; 29(4): 167–172. PubMed Abstract | Publisher Full Text
6. Basnaker M, Srikala SP, Satish BN: Cyclin d1 gene expression in oral mucosa of tobacco chewers–an immunohistochemical study. J. Clin. Diagn. Res. 2014 May; 8(5): ZC70–ZC75. PubMed Abstract | Publisher Full Text
7. Laco J, Mottl R, Höbling W, et al.: Cyclin D1 expression in ectomesenchymal chondromyxoid tumor of the anterior tongue. Int. J. Surg. Pathol. 2016 Oct; 24(7): 586–594. PubMed Abstract | Publisher Full Text
8. Miyamoto R, Uzawa N, Nagaoka S, et al.: Prognostic significance of cyclin D1 amplification and overexpression in oral squamous cell carcinomas. Oral Oncol. 2003 Sep 1; 39(6): 610–618. PubMed Abstract | Publisher Full Text
9. Ramos-García P, González-Moles MÁ, Ayén Á, et al.: Asymmetrical proliferative pattern loss linked to cyclin D1 overexpression in adjacent non-tumour epithelium in oral squamous cell carcinoma. Arch. Oral Biol. 2019 Jan 1; 97: 12–17. PubMed Abstract | Publisher Full Text
10. Bova RJ, Quinn DI, Nankervis JS, et al.: Cyclin D1 and p16INK4A expression predict reduced survival in carcinoma of the anterior tongue. Clin. Cancer Res. 1999 Oct; 5(10): 2810–2819. PubMed Abstract
11. Angadi PV, Krishnapillai R: Cyclin D1 expression in oral squamous cell carcinoma and verrucous carcinoma: correlation with histological differentiation. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. 2007 Mar 1; 103(3): e30–e35. PubMed Abstract | Publisher Full Text

Comments on this article Comments (0)

Version 1

VERSION 1 PUBLISHED 01 Sep 2023

Author details Author details

¹ “Department of Oral & Maxillofacial Pathology and Microbiology ”, Post graduate student, Sharad Pawar Dental College & Hospital, Datta Meghe Institute of Higher Education and Research, “Sawangi (Meghe)”, Wardha, Wardha, Maharashtra, 442004, India
² “Department of Oral & Maxillofacial Pathology and Microbiology”, “Sharad Pawar Dental College & Hospital”, Professor, “Datta Meghe Institute of Higher Education and Research , Sawangi (Meghe), Wardha, Wardha, Maharashtra, 442004, India

ANKITA CHAVHAN
Roles: Conceptualization, Formal Analysis, Funding Acquisition, Investigation, Methodology, Project Administration, Resources, Software, Supervision, Validation, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing

Swati Patil
Roles: Supervision, Validation, Visualization

Madhuri Gawande
Roles: Supervision, Validation, Visualization

Alka Hande
Roles: Supervision, Validation, Visualization

Archana Sonone
Roles: Supervision, Validation, Visualization

Aayushi Pakhale
Roles: Supervision, Validation, Visualization

Competing interests

No competing interests were disclosed.

Grant information

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

Article Versions (1)

version 1

Published: 01 Sep 2023, 12:1096

https://doi.org/10.12688/f1000research.134525.1

Copyright

© 2023 CHAVHAN A 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.

Download

Export To

metrics

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

Citations

0

SEE MORE DETAILS

CITE

how to cite this article

CHAVHAN A, Patil S, Gawande M et al. Protocol for the evaluation of cyclin D1 expression in tumor tissue and surgical margins in oral squamous cell carcinoma – a retrospective study [version 1; peer review: 1 not approved]. F1000Research 2023, 12:1096 (https://doi.org/10.12688/f1000research.134525.1)

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 1

VERSION 1

PUBLISHED 01 Sep 2023

Views

2

Reviewer Report 07 May 2024

KIRAN JADHAV, Department of Oral Pathology and Microbiology, Vasantdada Patil Dental College and Hospital, Budhgaon, Maharashtra, India; Oral Pathology and Microbiology, Vasant Dada Patil Dental College and Hospital, SANGLI, MAHARASHTRA, India

Not Approved

https://doi.org/10.5256/f1000research.147587.r253012

The major issues with article are as follows:

If it is a protocol then the aim and objectives, and material methodology should be written in future tense.
In the introduction the

The major issues with article are as follows:

If it is a protocol then the aim and objectives, and material methodology should be written in future tense.
In the introduction the authors should address the gap of knowledge in the literature so that the rationale of the study will be clear.
Discussion should not be just a literature review, the authors should justify results with reference to previously published literature.

Minor points:

The title of article is misleading and it does not match with the objectives of study.
The abstract can be rewritten to highlight the importance of the study, describing a succinct summary of the study.
The sample calculation references are not necessary to include in the manuscript text.

Is the rationale for, and objectives of, the study clearly described?

No
Is the study design appropriate for the research question?

No
Are sufficient details of the methods provided to allow replication by others?

No
Are the datasets clearly presented in a useable and accessible format?

Partly

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Oral and Maxillofacial Pathology, Oral Cancer, Lymph node metastasis in Oral cancer, micro RNA in Saliva, Oral exfoliative cytology

I confirm that I have read this submission and believe that I have an appropriate level of expertise to state that I do not consider it to be of an acceptable scientific standard, for reasons outlined above.

CITE

Report a concern

Respond or Comment

Comments on this article Comments (0)

Version 1

VERSION 1 PUBLISHED 01 Sep 2023

Open Peer Review

Reviewer Status

Reviewer Reports

	Invited Reviewers
	1
Version 1 01 Sep 23	read

KIRAN JADHAV, Vasantdada Patil Dental College and Hospital, Budhgaon, India; Vasant Dada Patil Dental College and Hospital, SANGLI, India

Comments on this article

All Comments(0)

Add a comment

Sign up for content alerts

Browse by related subjects

Back to all reports

Reviewer Report

2 Views

07 May 2024 | for Version 1

KIRAN JADHAV, Department of Oral Pathology and Microbiology, Vasantdada Patil Dental College and Hospital, Budhgaon, Maharashtra, India; Oral Pathology and Microbiology, Vasant Dada Patil Dental College and Hospital, SANGLI, MAHARASHTRA, India

2 Views Cite this report Responses(0)

Not Approved

The major issues with article are as follows:

If it is a protocol then the aim and objectives, and material methodology should be written in future tense.
In the introduction the authors should address the gap of knowledge in the literature so that the rationale of the study will be clear.
Discussion should not be just a literature review, the authors should justify results with reference to previously published literature.

Minor points:

The title of article is misleading and it does not match with the objectives of study.
The abstract can be rewritten to highlight the importance of the study, describing a succinct summary of the study.
The sample calculation references are not necessary to include in the manuscript text.

Is the rationale for, and objectives of, the study clearly described?

No
Is the study design appropriate for the research question?

No
Are sufficient details of the methods provided to allow replication by others?

No
Are the datasets clearly presented in a useable and accessible format?

Partly

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Oral and Maxillofacial Pathology, Oral Cancer, Lymph node metastasis in Oral cancer, micro RNA in Saliva, Oral exfoliative cytology

I confirm that I have read this submission and believe that I have an appropriate level of expertise to state that I do not consider it to be of an acceptable scientific standard, for reasons outlined above.

Respond to this report

Responses (0)

[1] 1. Sherkheli MA, Gisselmann G, Hatt H: Supercooling agent icilin blocks a warmth-sensing ion channel TRPV3. Sci. World J. 2012 Jan 1; 2012: 1–8. PubMed Abstract | Publisher Full Text | Free Full Text

[2] 2. Ohnishi Y, Watanabe M, Wato M, et al.: Cyclin D1 expression is correlated with cell differentiation and cell proliferation in oral squamous cell carcinomas. Oncol. Lett. 2014 Apr 1; 7(4): 1123–1127. PubMed Abstract | Publisher Full Text | Free Full Text

[3] 3. Mishra R, Das BR: Cyclin D1 expression and its possible regulation in chewing tobacco mediated oral squamous cell carcinoma progression. Arch. Oral Biol. 2009 Oct 1; 54(10): 917–923. PubMed Abstract | Publisher Full Text

[4] 4. Moharil RB, Khandekar S, Dive A, et al.: Cyclin D1 in oral premalignant lesions and oral squamous cell carcinoma: An immunohistochemical study. Journal of Oral and Maxillofacial Pathology: JOMFP. 2020 May; 24(2): 397. PubMed Abstract | Publisher Full Text | Free Full Text

[5] 5. Lam KY, Ng IO, Yuen AP, et al.: Cyclin D1 expression in oral squamous cell carcinomas: clinicopathological relevance and correlation with p53 expression. J. Oral Pathol. Med. 2000 Apr; 29(4): 167–172. PubMed Abstract | Publisher Full Text

[6] 6. Basnaker M, Srikala SP, Satish BN: Cyclin d1 gene expression in oral mucosa of tobacco chewers–an immunohistochemical study. J. Clin. Diagn. Res. 2014 May; 8(5): ZC70–ZC75. PubMed Abstract | Publisher Full Text

[7] 7. Laco J, Mottl R, Höbling W, et al.: Cyclin D1 expression in ectomesenchymal chondromyxoid tumor of the anterior tongue. Int. J. Surg. Pathol. 2016 Oct; 24(7): 586–594. PubMed Abstract | Publisher Full Text

[8] 8. Miyamoto R, Uzawa N, Nagaoka S, et al.: Prognostic significance of cyclin D1 amplification and overexpression in oral squamous cell carcinomas. Oral Oncol. 2003 Sep 1; 39(6): 610–618. PubMed Abstract | Publisher Full Text

[9] 9. Ramos-García P, González-Moles MÁ, Ayén Á, et al.: Asymmetrical proliferative pattern loss linked to cyclin D1 overexpression in adjacent non-tumour epithelium in oral squamous cell carcinoma. Arch. Oral Biol. 2019 Jan 1; 97: 12–17. PubMed Abstract | Publisher Full Text

[10] 10. Bova RJ, Quinn DI, Nankervis JS, et al.: Cyclin D1 and p16INK4A expression predict reduced survival in carcinoma of the anterior tongue. Clin. Cancer Res. 1999 Oct; 5(10): 2810–2819. PubMed Abstract

[11] 11. Angadi PV, Krishnapillai R: Cyclin D1 expression in oral squamous cell carcinoma and verrucous carcinoma: correlation with histological differentiation. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. 2007 Mar 1; 103(3): e30–e35. PubMed Abstract | Publisher Full Text

Protocol for the evaluation of cyclin D1 expression in tumor tissue and surgical margins in oral squamous cell carcinoma – a retrospective study

Abstract

Keywords

Introduction

Background and rationale

Objectives

Methods

Study Setting

Eligibility Criteria

Intervention

Outcomes

Primary outcome

Secondary outcome

Sample Size

Immunostaining

Dissemination

Study status

Discussion

Ethical considerations

Data availability

Reporting guidelines

Acknowledgements

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

Competing Interests Policy

Stay Updated