Association between eNOS rs1799983 (894G&gt;T) Polymorphism with Cancer and Stroke Risk: A meta-analysis

Abdul Rohim Tualeka; Juliana Jalaludin; Janvier Gasana; Nor Ashikin Sopian; How Ran Chao; Mohd Yusmaidie; Velu Perumal; Suardi Zurimi; Pudji Rahmawati; Ahsan Ahsan; Salsabila Novianti

doi:10.12688/f1000research.134992.3

Home Browse Association between eNOS rs1799983 (894G>T) Polymorphism with Cancer...

ALL Metrics

-

Views

-

Downloads

Get PDF

Get XML

Export

▬

✚

Systematic Review

Revised

Association between eNOS rs1799983 (894G>T) Polymorphism with Cancer and Stroke Risk: A meta-analysis

[version 3; peer review: 1 approved with reservations, 3 not approved]

Previous Title 'Association between nitric oxide and cancer and stroke risk: A meta-analysis'

Abdul Rohim Tualeka ¹, Juliana Jalaludin², Janvier Gasana^1,3, [...] Nor Ashikin Sopian^1,4, How Ran Chao^1,5, Mohd Yusmaidie^1,6, Velu Perumal^1,7, Suardi Zurimi⁸, Pudji Rahmawati⁹, Ahsan Ahsan¹⁰, Salsabila Novianti¹¹

Abdul Rohim Tualeka ¹, Juliana Jalaludin², [...] Janvier Gasana^1,3, Nor Ashikin Sopian^1,4, How Ran Chao^1,5, Mohd Yusmaidie^1,6, Velu Perumal^1,7, Suardi Zurimi⁸, Pudji Rahmawati⁹, Ahsan Ahsan¹⁰, Salsabila Novianti¹¹

PUBLISHED 10 Jun 2024

Author details Author details

¹ Department of Occupational Health and Safety, Faculty of Public Health, Universitas Airlangga, Surabaya, East Java, 60115, Indonesia
² Department of Environmental and Occupational Health, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia
³ Department of Environmental & Occupational Health, College of Public Health, Health Sciences Center Kuwait University, Kuwait University, Kuwait City, Shadadiya, Kuwait
⁴ Occupational Safety and Health Program, Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, Kuantan, Pahang, 26300, Malaysia
⁵ Department of Environmental Science and Engineering, National Pingtung University of Science and Technology, Pingtung, Neipu, Taiwan
⁶ Department of Toxicology, Advanced Medical & Dental Institute, Universiti Sains Malaysia, Bertam Kepala Bartas, Penang, 13200, Malaysia
⁷ Department of Industrial Design, Faculty of Design and Architecture, Universiti Putra Malaysia, Serdang, Selangor, 43400, Malaysia
⁸ Politeknik Kesehatan Kemenkes Maluku, Maluku, Indonesia
⁹ Department of Development of Islamic Society, Universitas Islam Negeri Sunan Ampel Surabaya, Surabaya, East Java, Indonesia
¹⁰ Faculty of Nurse, Universitas Brawijaya, Malang, East Java, Indonesia
¹¹ Department of Environmental Health, Faculty of Public Health, Universitas Airlangga, Surabaya, East Java, 60115, Indonesia

Abdul Rohim Tualeka
Roles: Conceptualization, Funding Acquisition, Methodology, Project Administration, Resources, Validation, Writing – Original Draft Preparation, Writing – Review & Editing

Juliana Jalaludin
Roles: Writing – Review & Editing

Janvier Gasana
Roles: Writing – Review & Editing

Nor Ashikin Sopian
Roles: Writing – Review & Editing

How Ran Chao
Roles: Writing – Review & Editing

Mohd Yusmaidie
Roles: Writing – Review & Editing

Velu Perumal
Roles: Supervision, Validation, Writing – Original Draft Preparation

Suardi Zurimi
Roles: Writing – Review & Editing

Pudji Rahmawati
Roles: Writing – Review & Editing

Ahsan Ahsan
Roles: Writing – Review & Editing

Salsabila Novianti
Roles: Project Administration, Writing – Original Draft Preparation

OPEN PEER REVIEW

REVIEWER STATUS

This article is included in the Oncology gateway.

Abstract

Background

Although numerous case-control studies have examined the role of nitric oxide, particularly the 894G>T polymorphism in the eNOS gene, in increasing the risk of stroke and cancer, there remains a need for a comprehensive meta-analysis to clarify these associations. This study aims to address this gap by thoroughly evaluating the relationship between the eNOS 894G>T polymorphism and the risks of cancer and stroke.

Methods

We conducted an exhaustive search across digital databases including Science Direct, PubMed, and Google Scholar for studies published between 2012-2023. A rigorous selection process was employed to include relevant studies, which were then analyzed using robust meta-analytical techniques to determine the association between the eNOS 894G>T polymorphism and the risks of cancer and stroke.

Results

In this meta-analysis, we combined data from 2,013 cases and 2,187 control subjects for cancer risk assessment and 1,006 cases with 1,146 control subjects for stroke risk evaluation. Our findings indicate that the eNOS 894G>T polymorphism is significantly associated with an increased risk of cancer when comparing GG vs. GT+TT genotypes. Additionally, there is a notable correlation between this polymorphism and stroke incidence under various genetic models (T vs. G, TT vs. GG + GT, GG + GT vs. TT).

Conclusions

The results of this meta-analysis suggest a significant association between the eNOS 894G>T polymorphism and increased risks of cancer and stroke. These findings underscore the importance of conducting future studies with larger sample sizes and more comprehensive analyses further to elucidate the role of nitric oxide in these diseases. This study addresses some concerns but further detailed and non-repetitive research is necessary for conclusive evidence.

Keywords

Nitric oxide, eNOS 894G>T, polymorphism, cancer, stroke, meta-analysis, safe work

Corresponding author: Abdul Rohim Tualeka

Competing interests: No competing interests were disclosed.

Grant information: This research was funded by Lembaga Penelitian dan Pengabdian Kepada Masyarakat Universitas Airlangga, grant number 333/UN3.15/PT/2023.
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Copyright: © 2024 Tualeka AR 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: Tualeka AR, Jalaludin J, Gasana J et al. Association between eNOS rs1799983 (894G>T) Polymorphism with Cancer and Stroke Risk: A meta-analysis [version 3; peer review: 1 approved with reservations, 3 not approved]. F1000Research 2024, 12:1467 (https://doi.org/10.12688/f1000research.134992.3) First published: 13 Nov 2023, 12:1467 (https://doi.org/10.12688/f1000research.134992.1) Latest published: 10 Jun 2024, 12:1467 (https://doi.org/10.12688/f1000research.134992.3)

Revised Amendments from Version 2

We have endeavored to reduce repetition, deepen analyses, and provide more meaningful insights in line with reviewers' comments. Titles have been corrected to include specific genes and polymorphisms for clarity. The study objectives have been appropriately placed in the Background section instead of the Methods section. The article has also provided clearer and more detailed information regarding the number of cases and controls, as per the reviewers' feedback.

To read any peer review reports and author responses for this article, follow the "read" links in the Open Peer Review table.

Introduction

Nitric oxide (NO) is a critical signaling molecule involved in various physiological and pathological processes in mammals. In humans, NO is produced enzymatically by nitric oxide synthase (NOS) from L-arginine, resulting in the formation of L-citrulline and NO itself (Korde Choudhari et al., 2013). The NOS family comprises three isoforms: endothelial NOS (eNOS), neuronal NOS (nNOS), and inducible NOS (iNOS) (Sachdev, 1999). NO plays a significant role in coagulation, neuronal activity, and cerebral blood flow regulation (Korde Choudhari et al., 2013). Moreover, it can induce cellular inflammation, potentially delaying stroke onset, and acting as a carcinogen, thereby increasing cancer risk.

Recent studies have highlighted the role of NO in the progression and metastasis of cancer through various mechanisms, including polyamine synthesis and inhibition of NO-mediated tumor cytotoxicity (Gào and Schöttker, 2017; Gào et al., 2019). NO’s involvement in hypoxic responses further promotes angiogenesis and cancer cell defense, which are attributed to its mutagenic properties. Specifically, iNOS has been implicated in carcinogenesis due to its production during chronic inflammation (Utispan and Koontongkaew, 2020).

The endothelial NOS (eNOS) isoform is vital for maintaining normal vascular tone under physiological conditions and has been extensively studied in the context of carcinogenesis and neuroprotection during stroke (Lim et al., 2008). Among the single nucleotide polymorphisms (SNPs) identified in the eNOS gene, the 894G>T (rs1799983) variant has been widely associated with cardiovascular diseases and cancer (Yang et al., 2019). This SNP results in structural and functional changes in the eNOS protein, contributing to the pathogenesis of these diseases.

Despite the recognition of multiple SNPs in the eNOS gene, this study focuses on the 894G>T variant due to its significant association with cancer and stroke risks. This meta-analysis aims to provide a comprehensive understanding of the relationship between eNOS 894G>T polymorphism and the risks of developing cancer and stroke.

Methods

Search strategy

The research method used was a meta-analysis. Our meta-analysis adhered to the criteria recommended in the Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines, or PRISMA. To discover relevant primary articles, we performed a comprehensive search of digital databases, including PubMed, Science Direct, and Google Scholar, to identify all relevant studies on the correlation of NO especially eNOS 894>GT, and risk to cancer and stroke. Articles sought must be published between 2012-2023. We applied the following keywords: “NO” or “NOS” or “eNOS” or “eNOS 894G>T” AND “polymorphism” AND “cancer risk” AND “stroke risk”. To ensure a comprehensive review of the literature, we conducted a thorough examination of the reference lists included in the recognized literature.

Inclusion and exclusion standard

Determination of inclusion and exclusion criteria followed PICOS (Problem, Intervention, Comparison, Outcome, and Study design). The included studies were carefully examined to ensure their relevance and quality for our study. No national restrictions were imposed, meaning studies from all countries were considered eligible for inclusion. The research that met the eligibility criteria was carefully selected for our analysis: 1) Articles published in the English language that investigated the correlation between NO, especially eNOS 894G>T, and the risk of cancer and stroke; 2) Designed as a case-control study; 3) Articles that provided detailed data on genotype and allele frequencies of eNOS gene polymorphisms, which has sufficient data for the calculation of the odds ratio (OR) and the confidence interval of 95%. Therefore, studies were excluded according to as the following criteria: 1) Qualitative research; 2) No available genotype frequency; 3) Studies without control; 4) Meta-analysis studies; and 5) Animal studies.

Data extraction

The data in all studies were extracted when sufficient criteria were met. We then used Microsoft Excel to record the year of publication, the last name of the authors, control, and case sample sizes, and the country of the study. The results were then compared after being extracted, and an assessment was carried out along with the resolution of matters that were not appropriate through consensus. We extracted data from the nine articles meeting eligibility criteria for cancer risk and the seven articles for stroke risk association with NO.

Statistical analysis

A statistical review was implemented using RevMan, Cochrane with version 5.4 to investigate the association between NO and the risk of stroke and cancer. Crude ORs and a CI of 95% were utilized. Pooled ORs were computed for various genetic models of the eNOS G894T gene polymorphism, including GT+TT versus GG, GT versus GG, TT versus GG, and T versus G. The eNOS gene encodes for endothelial nitric oxide synthase and has a polymorphism at position 894G>T that can result in GG, GT, or TT variants (Buldreghini et al., 2010). G represents the homozygous wild-type genotype, where the individual has two copies of the G allele. GT represents the heterozygous genotype, where the individual has one copy of the G allele and one copy of the T allele. TT represents the homozygous variant genotype, where the individual has two copies of the T allele (Hinz et al., 2013). The calculation of pooled ORs allowed to perform a Z test with a significance level of p≤0.05.

The presence of heterogeneity among the studies included was assessed with a Q test score. If there was no significant hterogeneity, i.e., p>0.10, the effect model was applied consistently. Otherwise, when (p<0.10), the random-effects model was utilized. The diversity of the included research was assessed using the I² test, which quantifies the degree of heterogeneity. If the I² value was less than 25%, it indicated no heterogeneity. If the I² value ranged from 25% to 50%, it showed moderate heterogeneity. If the I² was greater than 50%, it indicated extreme heterogeneity. The 50% p-value indicating the existence of heterogeneity between the studies and a random effects model (Mantel-Haenszel technique) was implemented; conversely, if no significant heterogeneity was found, the fixed effect model is applied.

Results

Study processing

The flow diagram in Figure 1 summarises the study workflow. A total of 145 articles were identified in the databases. After removing duplicates, a total of 109 studies remained. 36 studies were screened, and 21 studies were excluded for various reasons. Ten other studies (da Costa Escobar Piccoli et al., 2012; Jang et al., 2013; Rah et al., 2013; Akhter et al., 2014; Kang et al., 2014; Özçelik et al., 2014; Ben Chaaben et al., 2015; Hung et al., 2019; Lee, 2019; Tsay et al., 2019) were also excluded because of unavailable genotype frequency. Six studies (Hao, Montiel and Huang, 2010; Yao et al., 2013; Guo, 2014; Zhao et al., 2014; Abedinzadeh et al., 2020; Akbar et al., 2022) were not included in the analysis because they were meta-analysis studies.

Figure 1. Flow diagram depicting the inclusion process of the studies in the meta-analysis.

Finally, fifteen studies met the eligibility criteria and were included in the meta-analysis. These consisted of nine case-control studies examining the association between nitric oxide (NO) and cancer risk, and six studies analyzing the association between NO and stroke risk. The characteristics of these studies are summarized in Table 1.

Table 1. Characteristics and genetic frequencies derived the studies included in the meta-analysis.

First Author/Year	Country	Risk	Case/Control	Case					Control
				Genotype			Allele		Genotype			Allele
				GG	TT	GT	T	G	GG	TT	GT	T	G
(Adibmanesh et al., 2020)	Iran	Cancer	100/100	28	28	44	100	100	57	6	37	49	151
(Aouf et al., 2019)	Tunisia	Cancer	259/169	149	20	90	130	388	73	18	78	114	224
(Branković et al., 2013)	Serbia	Cancer	150/100	76	9	65	83	217	54	6	40	52	148
(Carkic et al., 2020)	Serbia	Cancer	50/110	21	5	24	34	66	61	7	42	56	164
(Koçer et al., 2020)	Turkey	Cancer	107/100	74	1	32	34	180	65	1	34	36	164
(Su et al., 2018)	Taiwan	Cancer	1044/1200	825	10	209	229	1859	935	15	250	280	2120
(Verim et al., 2013)	Turkey	Cancer	66/88	7	10	49	69	63	31	13	44	70	106
(Yadav et al., 2019)	India	Cancer	179/173	88	20	64	104	240	96	8	59	75	251
(Yanar et al., 2016)	Turkey	Cancer	58/147	18	11	29	51	65	31	35	81	151	143
(Anliaçik et al., 2019)	Turkey	Stroke	112/160	21	14	77	44	40	38	19	103	57	61
(Diakite et al., 2014)	Morocco	Stroke	165/182	83	16	66	30	70	117	7	58	20	81
(El Gohary, El Azab and Kamal El-Din, 2017)	Egypt	Stroke	30/10	18	6	6	45	15	5	3	3	12	8
(Kaur, Uppal and Kaur, 2015)	India	Stroke	120/101	84	6	30	18	83	83	1	17	9	91
(Kumar et al., 2016)	India	Stroke	250/250	164	12	74	20	80	186	5	59	14	86
(Shyu et al., 2017)	Taiwan	Stroke	229/243	151	16	62	21	50	185	7	51	13	87

Meta-analysis

The meta-analysis incorporated data from a total of 2,013 cases and 2,187 control subjects for cancer risk assessment, and 1,006 cases with 1,146 control subjects for stroke risk evaluation. Table 2 presents the aggregated outcome polymorphism through meta-analysis and its association with cancer risk. The results showed that the genetic model “GT+TT vs GG” showed a significant association with cancer risk, as it had a significant p-value (<0.05) and a high OR (1.96). Other genetic models did not show a significant association with cancer risk, indicating that specific genotype combinations might play a more crucial role in cancer susceptibility.

Table 2. The relationship of cancer risk with eNOS G894T polymorphism in summary.

Genetic Models	NS	Pooled ORs (95% CI)	p-value^a (Z test)	I² (%)	pH	pE	Method
T vs G	9	1.00 (0.44,2.27)	1.00	94	<0.00001	0.17497	Random model
G vs T	9	1.00 (0.44,2.27)	1.00	94	<0.00001	0.34809	Random model
GT+TT vs GG	9	1.96 (1.22,3.15)	0.005	85	<0.00001	0.42245	Random model
TT vs GG+GT	9	0.51 (0.22-1.17)	0.11	84	<0.00001	0.03886	Random model
GT vs GG+TT	9	1.21 (0.77-1.91)	0.41	84	<0.00001	0.13326	Random model

a The p-value of the Z test was found to be less than 0.05.

Frequencies of T versus G, GT+TT versus GG, GT versus GG+TT, and TT versus GG+GT genotypes were 78% versus 78%, 58% versus 31%, 48% versus 33%, and 62% versus 38%, respectively in cases and controls.

Table 3 analysis revealed that individuals with the T allele (either in GT or TT genotypes) have a higher risk of stroke compared to those with the G allele (p < 0.05). This supports the hypothesis that the eNOS 894G>T polymorphism may be a significant factor in stroke risk.

Table 3. The relationship of stroke risk with eNOS G894T polymorphism in summary.

Genetic models	NS	Pooled ORs (95% CI)	p-value^a (Z test)	I² (%)	pH	pE	Method
T versus G	6	1.20 (1.01,1.43)	0.04	45	0.11	0.11583	Fixed model
G versus T	6	0.88 (0.74,1.05)	0.15	0	0.86	0.17541	Fixed model
GT+TT versus GG	6	0.68 (0.24-1.93)	0.47	91	<0.00001	0.17523	Random model
TT versus GG+GT	6	0.09 (0.03-0.30)	0.0001	94	<0.00001	0.44879	Random model
GT versus GG+TT	6	1.03 (0.40-2.64)	0.95	92	<0.00001	0.29785	Random model

a The p-value of the Z test was found to be less than 0.05, indicating statistical significance.

Frequencies of T versus G, GT versus GG+TT, GT+TT versus GG, and TT versus GG+GT genotypes were 37% versus 27%, 76% versus 74%, 78% versus 81%, and 29% versus 84%, respectively in cases and controls.

Heterogeneity among studies

Heterogeneity was observed among the studies in every allele and gene, as depicted in Figures 2 and 3 (T versus G, G versus T, GT+TT versus GG, TT versus GG + GT, and GT versus GG + TT). Tables 2 and 3 provide information on the selected model (random or fixed effect) utilized in order to review universal genetic model correlations.

Figure 2. Forest plots depicting the association of eNOS 894G>T polymorphisms with cancer risk in all genetic models, including: A). T vs G, B). GT + TT vs GG, C). TT vs GG + GT, and D). GT vs GG + TT, respectively.

Figure 3. Forest plots illustrating the association of eNOS 894G>T polymorphisms with stroke risk in all genetic models, including: A). T vs G, B). GT + TT vs GG, C). TT vs GG + GT, and D). GT vs GG + TT, respectively.

Publication bias

Both funnel plot images (Figure 4) show the possibility of publication bias. The left side of the plot is more “full” than the right side (asymmetry of the funnel plot). This indicates that there are more studies with positive results (OR>1) compared to studies with negative results (OR<1). Egger’s test results show a p-value<0.05, which means there is statistical evidence that the funnel plot is significantly asymmetrical.

Figure 4. Funnel Plot of association between eNOS 894G>T polymorphism and risk of cancer and stroke.

Discussion

NO acts as a crucial part of numerous pathological and psychological processes, including cancer development and stroke (Zhao et al., 2014). This meta-analysis aimed to elucidate the association between the eNOS 894G>T polymorphism and the risk of cancer and stroke. Our findings highlight several important points that contribute to the current understanding of NO’s involvement in these diseases. Our findings suggest a potential link between the polymorphism and increased cancer susceptibility, particularly in individuals with the TT or GT genotypes compared to GG. This association appears consistent across various ethnicities (African, Asian, European). However, some studies reported no significant association, highlighting the potential influence of other factors. Similarly, the eNOS 894G>T polymorphism might be associated with an increased risk of stroke, especially in the “T versus G” and “TT versus GG+GT” genetic models. However, findings here were more varied, with some studies showing no significant association.

Figure 5 and Figure 6 depict the potential roles of NO in cancer and stroke development. Reduced eNOS activity and subsequent NO deficiency might contribute to tumor growth and impaired blood flow in stroke. External factors like chronic stress, known to affect eNOS activity, could further influence cancer and stroke risks. Based on Figure 5, overproduction of NO can facilitate tumor angiogenesis and metastasis.

Figure 5. The role of nitric oxide in cancer.

Based on Figure 6, low levels of NO derived from eNOS may exert neuroprotection in stroke by promoting vasodilatation and increasing cerebral blood flow (Yang et al., 2019). However, at the same time, there is an enhancement in superoxide production due to eNOS uncoupling. When eNOS is uncoupled, nitric oxide (NO) is not produced, and peroxynitrite is formed instead. This occurs when the enzyme is unable to convert L-arginine into NO due to a lack of cofactors or substrates. Peroxynitrite damages lipids, proteins, and DNA and can trigger the activation of polyadenosine diphosphate ribose (ADP-ribose) polymerase (PARP), all of which contribute to neurotoxicity in stroke.

Figure 6. The role of nitric oxide in stroke.

However, this study has several limitations that need to be considered. Firstly, significant heterogeneity between the studies included in the analysis may affect the validity and generalisability of the results. Furthermore, interpretation of the results should be done with caution as additional uncontrolled factors, such as differences in the sample population, and environment, may affect the estimation of genetic effects. Furthermore, in some cases, the limited number of studies may have limited the statistical power of the analyses and affected the ability to draw strong conclusions. Therefore, more extensive and well-controlled studies are needed to confirm these findings and understand more about the role of genetics in the pathogenesis of stroke and cancer diseases.

Conclusions

In conclusion, the recent meta-analysis found that nitric oxide-related polymorphisms with the eNOS 894G>T gene are associated with a substantial risk of cancer in the total population based on the GG vs. GT+TT genetic model and significantly correlated with the manifestation of stroke in the genetic models T vs. G, TT vs. GG + GT, and GG + GT vs. TT. Considering the conclusion, these results should be reassessed in the coming days through studies with a larger sample population.

Data availability

Underlying data

All underlying data are available as part of the article and no additional source data are required.

Reporting guidelines

Zenodo: PRISMA Checklist for “Association between nitric oxide and cancer and stroke risk: A meta-analysis”, https://doi.org/10.5281/zenodo.8031323 (Tualeka et al., 2023).

Data are available under the terms of the Creative Commons Zero “No rights reserved” data waiver (CC0 1.0 Public domain dedication).

References

Abedinzadeh M, Dastgheib SA, Maleki H, et al.: Association of endothelial nitric oxide synthase gene polymorphisms with susceptibility to prostate cancer: A comprehensive systematic review and meta-analysis. Urol. J. 2020; 17(4): 329–337. PubMed Abstract | Publisher Full Text
Adibmanesh A, Bijanzadeh M, Mohammadzadeh G, et al.: The correlation of endothelial nitric oxide synthase gene rs1799983 polymorphisms with colorectal cancer. Int. J. Cancer Manag. 2020; 13(5). Publisher Full Text
Akbar KR, Permatasari AI, Suwarno B, et al.: Lack of association between endothelial nitric oxide synthase (eNOS) G894T gene polymorphism and the risk of bladder cancer Α meta-analysis. Arch. Hell. Med. 2022; 39(5): 647–653.
Akhter MS, Biswas A, Rashid H, et al.: Screening of the NOS3 gene identifies the variants 894G/T, 1998C/G and 2479G/A to be associated with acute onset ischemic stroke in young Asian Indians. J. Neurol. Sci. 2014; 344(1–2): 69–75. PubMed Abstract | Publisher Full Text
Anliaçik SÖ, Tokgöz S, Zamani AG, et al.: Investigation of the relationship between ischemic stroke and endothelial nitric oxide synthase gene polymorphisms [G894T, intron 4 VNTR and T786C]. Turk. J. Med. Sci. 2019; 49(2): 589–594. PubMed Abstract | Publisher Full Text | Free Full Text
Aouf S, Laribi A, Gabbouj S, et al.: Contribution of Nitric oxide synthase 3 genetic variants to nasopharyngeal carcinoma risk and progression in a Tunisian population. Eur. Arch. Otorhinolaryngol. 2019; 276(4): 1231–1239. Publisher Full Text
Branković A, Brajušković G, Nikolić Z, et al.: Endothelial nitric oxide synthase gene polymorphisms and prostate cancer risk in serbian population. Int. J. Exp. Pathol. 2013; 94(6): 355–361. PubMed Abstract | Publisher Full Text | Free Full Text
Buldreghini E, Mahfouz RZ, Vignini A, et al.: Single Nucleotide Polymorphism (SNP) of the Endothelial Nitric Oxide Synthase (eNOS) Gene (Glu298Asp Variant) in Infertile Men With Asthenozoospermia. J. Androl. 2010; 31(5): 482–488. PubMed Abstract | Publisher Full Text
Carkic J, Nikolic N, Nisevic J, et al.: Endothelial nitric oxide synthase polymorphisms/haplotypes are strong modulators of oral cancer risk in serbian population. J. Oral Sci. 2020; 62(3): 322–326. PubMed Abstract | Publisher Full Text
Ben Chaaben A, Mariaselvam C, Salah S, et al.: Polymorphisms in oxidative stress-related genes are associated with nasopharyngeal carcinoma susceptibility. Immunobiology. 2015; 220(1): 20–25. Publisher Full Text
da Costa Escobar Piccoli J , Manfredini V, Hamester FI, et al.: Interaction Between Endothelial Nitric Oxide Synthase Gene Polymorphisms (-786T>C, 894G>T and Intron 4 a/b) and Cardiovascular Risk Factors in Acute Coronary Syndromes. Arch. Med. Res. 2012; 43(3): 205–211. PubMed Abstract | Publisher Full Text
Diakite B, Hamzi K, Slassi I, et al.: G894T endothelial nitric oxide synthase polymorphism and ischemic stroke in Morocco. Meta Gene. 2014; 2(1): 349–357. PubMed Abstract | Publisher Full Text | Free Full Text
Gào X, Schöttker B: Reduction-oxidation pathways involved in cancer development: A systematic review of literature reviews. Oncotarget. 2017; 8(31): 51888–51906. PubMed Abstract | Publisher Full Text | Free Full Text
Gào X, Xuan Y, Benner A, et al.: Nitric Oxide Metabolites and Lung Cancer Incidence: A Matched Case-Control Study Nested in the ESTHER Cohort. Oxidative Med. Cell. Longev. 2019; 2019: 1–9. PubMed Abstract | Publisher Full Text | Free Full Text
El Gohary EA, El Azab AA, Kamal El-Din MMM: Study of Relationship between G894T Variant of Endothelial Nitric Oxide Synthase Gene and Acute Ischemic Stroke. Egyptian J. Hosp. Med. 2017; 69(1): 1607–1613. Publisher Full Text
Guo X: Endothelial nitric oxide (eNOS) gene G894T and VNTR polymorphisms are closely associated with the risk of ischemic stroke development for Asians: Meta-analysis of epidemiological studies. Mol. Biol. Rep. 2014; 41(4): 2571–2583. PubMed Abstract | Publisher Full Text
Hao Y, Montiel R, Huang Y: Endothelial nitric oxide synthase (eNOS) 894 G>T polymorphism is associated with breast cancer risk: A meta-analysis. Breast Cancer Res. Treat. 2010; 124(3): 809–813. PubMed Abstract | Publisher Full Text
Hinz J, Schöndorf D, Bireta C, et al.: The eNOS 894G/T gene polymorphism and its influence on early and long-term mortality after on-pump cardiac surgery. J. Cardiothorac. Surg. 2013; 8(1): 199. PubMed Abstract | Publisher Full Text | Free Full Text
Hung WC, Wu TF, Ng SC, et al.: Involvement of endothelial nitric oxide synthase gene variants in the aggressiveness of uterine cervical cancer. J. Cancer. 2019; 10(12): 2594–2600. PubMed Abstract | Publisher Full Text | Free Full Text
Jang MJ, Jeon YJ, Kim JW, et al.: Association of eNOS polymorphisms (-786T>C, 4a4b, 894G>T) with colorectal cancer susceptibility in the Korean population. Gene. 2013; 512(2): 275–281. PubMed Abstract | Publisher Full Text
Kang MK, Kim OJ, Jeon YJ, et al.: Interplay between polymorphisms in the endothelial nitric oxide synthase (eNOS) gene and metabolic syndrome in determining the risk of ischemic stroke in Koreans. J. Neurol. Sci. 2014; 344(1–2): 55–59. PubMed Abstract | Publisher Full Text
Kaur K, Uppal A, Kaur A: An exonic G894T variant of endothelial nitric oxide synthase gene as a risk factor for ischemic stroke in North Indians. Acta Neurobiol. Exp. 2015; 75(4): 339–350. PubMed Abstract
Koçer C, Benlier N, Balci SO, et al.: The role of endothelial nitric oxide synthase gene polymorphisms in patients with lung cancer. Clin. Respir. J. 2020; 14(10): 948–955. PubMed Abstract | Publisher Full Text
Korde Choudhari S, Chaudhary M, Bagde S, et al.: Nitric oxide and cancer: A review. World J. Surg. Oncol. 2013; 11: 1–11. PubMed Abstract | Publisher Full Text | Free Full Text
Kumar A, Misra S, Kumar P, et al.: Association between Endothelial nitric oxide synthase G894T gene polymorphism and risk of ischemic stroke in North Indian population: A case-control study. Neurol. Res. 2016; 38(7): 575–579. PubMed Abstract | Publisher Full Text
Lee P-C: A Case-Control Study and Meta-Analysis of the Association of eNOS rs1799983 SNP with Stroke Risk. Med. Health. 2019; 14(1): 118–134. Publisher Full Text
Lim KH, Ancrile BB, Kashatus DF, et al.: Tumour maintenance is mediated by eNOS. Nature. 2008; 452(7187): 646–649. PubMed Abstract | Publisher Full Text | Free Full Text
Özçelik AT, Can Demirdöʇen B, Demirkaya Ş, et al.: Importance of NOS3 genetic polymorphisms in the risk of development of ischemic stroke in the Turkish population. Genet. Test. Mol. Biomarkers. 2014; 18(12): 797–803. PubMed Abstract | Publisher Full Text
Rah H, Jeon YJ, Lee WS, et al.: Association of nitric oxide synthase gene polymorphisms (-786T>C, 4a4b, 894G>T) with primary ovarian insufficiency in Korean women. Maturitas. 2013; 74(2): 160–165. PubMed Abstract | Publisher Full Text
Sachdev K: Clinical Pathology and Clinical Bacteriology. J. Clin. Pathol. Bacteriol. 1999; 22(1). Publisher Full Text
Shyu HY, Chen MH, Hsieh YH, et al.: Association of eNOS and Cav-1 gene polymorphisms with susceptibility risk of large artery atherosclerotic stroke. PLoS One. 2017; 12(3): e0174110–e0174112. PubMed Abstract | Publisher Full Text | Free Full Text
Su CW, Chien MH, Lin CW, et al.: Associations of genetic variations of the endothelial nitric oxide synthase gene and environmental carcinogens with oral cancer susceptibility and development. Nitric Oxide. 2018; 79(May): 1–7. PubMed Abstract | Publisher Full Text
Tsay MD, Hsieh MJ, Wang SS, et al.: Impact of endothelial nitric oxide synthase polymorphisms on urothelial cell carcinoma development. Urol. Oncol. 2019; 37(4): 293.e1–293.e9. PubMed Abstract | Publisher Full Text
Tualeka AR, Jalaludin J, Gasana J, et al.: PRISMA Checklist for Article Association between nitric oxide and cancer and stroke risk: A meta-analysis. Zenodo. 2023. Publisher Full Text
Utispan K, Koontongkaew S: High nitric oxide adaptation in isogenic primary and metastatic head and neck cancer cells. Anticancer Res. 2020; 40(5): 2657–2665. Publisher Full Text
Verim L, Toptaş B, Özkan NE, et al.: Possible relation between the NOS3 gene GLU298ASP polymorphism and bladder cancer in Turkey. Asian Pac. J. Cancer Prev. 2013; 14(2): 665–668. PubMed Abstract | Publisher Full Text
Yadav SK, Gupta S, Yadav A, et al.: Endothelial nitric oxide synthase gene polymorphisms modulate the risk of squamous cell carcinoma of head and neck in north Indian population. Meta Gene. 2019; 21(October 2018): 100575. Publisher Full Text
Yanar K, Çakatay U, Aydin S, et al.: Relation between Endothelial Nitric Oxide Synthase Genotypes and Oxidative Stress Markers in Larynx Cancer. Oxidative Med. Cell. Longev. 2016; 2016: 1–8. PubMed Abstract | Publisher Full Text | Free Full Text
Yang C, Hawkins KE, Doré S, et al.: Neuroinflammatory mechanisms of blood-brain barrier damage in ischemic stroke.2019.
Yao YS, Chang WW, Jin YL, et al.: An updated meta-analysis of endothelial nitric oxide synthase gene: Three well-characterized polymorphisms with ischemic stroke. Gene. 2013; 528(2): 84–92. PubMed Abstract | Publisher Full Text
Zhao C, Yan W, Zu X, et al.: Association between endothelial nitric oxide synthase 894G>T polymorphism and prostate cancer risk: a meta-analysis of literature studies. Tumor Biol. 2014; 35(12): 11727–11733. PubMed Abstract | Publisher Full Text

Comments on this article Comments (0)

Version 3

VERSION 3 PUBLISHED 13 Nov 2023

Author details Author details

¹ Department of Occupational Health and Safety, Faculty of Public Health, Universitas Airlangga, Surabaya, East Java, 60115, Indonesia
² Department of Environmental and Occupational Health, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia
³ Department of Environmental & Occupational Health, College of Public Health, Health Sciences Center Kuwait University, Kuwait University, Kuwait City, Shadadiya, Kuwait
⁴ Occupational Safety and Health Program, Faculty of Industrial Sciences and Technology, Universiti Malaysia Pahang, Kuantan, Pahang, 26300, Malaysia
⁵ Department of Environmental Science and Engineering, National Pingtung University of Science and Technology, Pingtung, Neipu, Taiwan
⁶ Department of Toxicology, Advanced Medical & Dental Institute, Universiti Sains Malaysia, Bertam Kepala Bartas, Penang, 13200, Malaysia
⁷ Department of Industrial Design, Faculty of Design and Architecture, Universiti Putra Malaysia, Serdang, Selangor, 43400, Malaysia
⁸ Politeknik Kesehatan Kemenkes Maluku, Maluku, Indonesia
⁹ Department of Development of Islamic Society, Universitas Islam Negeri Sunan Ampel Surabaya, Surabaya, East Java, Indonesia
¹⁰ Faculty of Nurse, Universitas Brawijaya, Malang, East Java, Indonesia
¹¹ Department of Environmental Health, Faculty of Public Health, Universitas Airlangga, Surabaya, East Java, 60115, Indonesia

Abdul Rohim Tualeka
Roles: Conceptualization, Funding Acquisition, Methodology, Project Administration, Resources, Validation, Writing – Original Draft Preparation, Writing – Review & Editing

Juliana Jalaludin
Roles: Writing – Review & Editing

Janvier Gasana
Roles: Writing – Review & Editing

Nor Ashikin Sopian
Roles: Writing – Review & Editing

How Ran Chao
Roles: Writing – Review & Editing

Mohd Yusmaidie
Roles: Writing – Review & Editing

Velu Perumal
Roles: Supervision, Validation, Writing – Original Draft Preparation

Suardi Zurimi
Roles: Writing – Review & Editing

Pudji Rahmawati
Roles: Writing – Review & Editing

Ahsan Ahsan
Roles: Writing – Review & Editing

Salsabila Novianti
Roles: Project Administration, Writing – Original Draft Preparation

Competing interests

No competing interests were disclosed.

Grant information

This research was funded by Lembaga Penelitian dan Pengabdian Kepada Masyarakat Universitas Airlangga, grant number 333/UN3.15/PT/2023.
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Article Versions (3)

version 3

Revised

Published: 10 Jun 2024, 12:1467

https://doi.org/10.12688/f1000research.134992.3

version 2

Revised

Published: 28 Mar 2024, 12:1467

https://doi.org/10.12688/f1000research.134992.2

version 1

Published: 13 Nov 2023, 12:1467

https://doi.org/10.12688/f1000research.134992.1

Copyright

© 2024 Tualeka AR 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

Tualeka AR, Jalaludin J, Gasana J et al. Association between eNOS rs1799983 (894G>T) Polymorphism with Cancer and Stroke Risk: A meta-analysis [version 3; peer review: 1 approved with reservations, 3 not approved]. F1000Research 2024, 12:1467 (https://doi.org/10.12688/f1000research.134992.3)

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

Current Reviewer Status: ?

Key to Reviewer Statuses VIEW HIDE

ApprovedThe 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 approvedFundamental flaws in the paper seriously undermine the findings and conclusions

Version 3

VERSION 3

PUBLISHED 10 Jun 2024

Revised

Views

1

Reviewer Report 31 Oct 2024

Saori Furuta, Department of Cell & Cancer Biology, College of Medicine and Life Sciences, University of Toledo Health Science Campus, MetroHealth Medical Center, Case Western Reserve University School of Medicine, Toledo, Cleveland, USA

Not Approved

https://doi.org/10.5256/f1000research.167579.r333227

In the present study, Tualeka et al. explored the association of eNOS 894G>T (Asp298Glu) Polymorphism with elevated risks of cancer and stroke through meta-analysis of published studies. While the pathogenic relevance of this polymorphism to various diseases has been extensively ... Continue reading

In the present study, Tualeka et al. explored the association of eNOS 894G>T (Asp298Glu) Polymorphism with elevated risks of cancer and stroke through meta-analysis of published studies. While the pathogenic relevance of this polymorphism to various diseases has been extensively studied, its relevance to stroke and cancer has not been fully determined yet. Their results concluded that there is a significant correlation of this polymorphism to both stoke and cancer, suggesting the roles of this polymorphism as the susceptibility marker for both conditions. Despite the potential significance of the study findings, there are several caveats described below that make it the present form of the manuscript unsuitable for publication. Major revisions would be suggested to improve the quality of the manuscript.
Specific comments:

Please describe the biological consequence of this polymorphism. Several studies showed that this polymorphism reduces the basal level of NO production. If so, the proposed models in Fig. 5 based on the elevated NO production is not correct.
Please describe the nature of studies utilized in tables, including gender, age, disease condition, etc.
Please separate analyses based on cancer types (e.g., tissues and grades).
Please describe all the acronyms in the text, figures, and tables.

Are the rationale for, and objectives of, the Systematic Review clearly stated?

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

No
Is the statistical analysis and its interpretation appropriate?

I cannot comment. A qualified statistician is required.
Are the conclusions drawn adequately supported by the results presented in the review?

Partly
If this is a Living Systematic Review, is the ‘living’ method appropriate and is the search schedule clearly defined and justified? (‘Living Systematic Review’ or a variation of this term should be included in the title.)

No

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Breast cancer, lung cancer, cancer immunology, cancer metabolism, microbiome

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

Views

3

Reviewer Report 06 Aug 2024

Mirza Saqib Baig, Department of Biosciences and Biomedical Engineering, Indian institute of technology indore, Indore, India

Approved with Reservations

https://doi.org/10.5256/f1000research.167579.r298825

The article entitled "Association of eNOS rs1799983 (894G>T) Polymorphism with Cancer and Stroke Risk: A Meta-Analysis" has derived a meta-analysis study of the effect of different genotypic variants on the function of eNOS and its subsequent role in the risk ... Continue reading

The article entitled "Association of eNOS rs1799983 (894G>T) Polymorphism with Cancer and Stroke Risk: A Meta-Analysis" has derived a meta-analysis study of the effect of different genotypic variants on the function of eNOS and its subsequent role in the risk of Cancer and Stroke. Although the study shows the role of different genotypic variants of eNOS G894T in the outcome of the disease, a few corrections are needed:
1. As most of the genotypes showed heterogeneity in the study, further analysis of the genotypes by deriving different subgroups from the study (e.g., age, sex, ethnicity) can help in identifying the main causes associated with the disease.
2. As the funnel plot showed publication bias in the study, the authors should provide a discussion on the steps to be taken to reduce the publication bias in the study.
3. It is not clear from where the values of the total and the events were given, as the values does not match the individual genotype and allele values in the case and control provided in the Table 2. Author should provide the text in the discussion how the value of the event and total were provided in the table corresponding to each study. Also, how the weight was assigned in the individual study in Figure 2 and 3.
5. The author should provide short discussion on the individual terms such as I², pH, pE etc and why it was used in the Table 2 and 3
6. Author should review the Figure 5 and 6 as the text provided in the discussion doesn’t correlate with the actual figure. Also, it is not clear in Figure 6, whether variant of eNOS leads to more uncoupling of the protein and the disease phenotype compared to control.
7. The author should provide more references of the actual in-vitro studies done on the eNOS G894T variant in the risk of cancer and stroke. Also, there should be more discussion in the text by which mechanism the eNOS G894T variant shows the disease phenotype in cancer and stroke.

Are the rationale for, and objectives of, the Systematic Review clearly stated?

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

Yes
Is the statistical analysis and its interpretation appropriate?

Yes
Are the conclusions drawn adequately supported by the results presented in the review?

No

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, however I have significant reservations, as outlined above.

CITE

Report a concern

Respond or Comment

Version 2

VERSION 2

PUBLISHED 28 Mar 2024

Revised

Views

7

Reviewer Report 02 May 2024

Katrina Miranda, University of Arizona, Arizona, USA

Not Approved

https://doi.org/10.5256/f1000research.163290.r260355

The manuscript is highly repetitive, cursory in nature and does not provide any major insights. ... Continue reading

CITE

Report a concern

Respond or Comment

Version 1

VERSION 1

PUBLISHED 13 Nov 2023

Views

25

Reviewer Report 16 Feb 2024

Katrina Miranda, University of Arizona, Arizona, USA

Not Approved

https://doi.org/10.5256/f1000research.148090.r233438

The manuscript by Tualeka and colleagues provides a meta-analysis on the correlation of a polymorphism in the eNOS gene on cancer and stroke. The analysis itself is fine. However, presentation of the complex relationship of NO and cancer or stroke ... Continue reading

The manuscript by Tualeka and colleagues provides a meta-analysis on the correlation of a polymorphism in the eNOS gene on cancer and stroke. The analysis itself is fine. However, presentation of the complex relationship of NO and cancer or stroke is cursory. References include randomly chosen reviews rather than the original literature. It would be beneficial to include an expert on NO in preparation of a revision. For example, the levels of NO produced by the isoforms is not well presented. The polymorphism is also not described sufficiently. Lately, in Figure 5, if eNOs is uncoupled, where does the NO come from to produce peroxynitrite? In sum, more details are needed in this manuscript.

Are the rationale for, and objectives of, the Systematic Review clearly stated?

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

Yes
Is the statistical analysis and its interpretation appropriate?

Yes
Are the conclusions drawn adequately supported by the results presented in the review?

Partly

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: NO and cancer

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

Views

32

Reviewer Report 12 Feb 2024

Eric Tzyy Jiann Chong, Biotechnology Research Institute, Universiti Malaysia Sabah, Sabah, Malaysia

Not Approved

https://doi.org/10.5256/f1000research.148090.r233428

The manuscript by Tualeka et al. aims to associate the eNOS rs1799983 SNP with the risk of cancer and stroke using a meta-analysis approach. The authors concluded that this SNP is associated with the risk of cancer and stroke in ... Continue reading

The manuscript by Tualeka et al. aims to associate the eNOS rs1799983 SNP with the risk of cancer and stroke using a meta-analysis approach. The authors concluded that this SNP is associated with the risk of cancer and stroke in some genetic model comparisons. Overall, the manuscript is poorly written, with many unclear statements and grammatical errors, although it has been reviewed and edited by many authors. There are many concerns that should be addressed by the authors.

Title
- The current title is not specific enough to reflect the contents of the manuscript. Please include the specific gene and polymorphism in the title.

Abstract
-Why is the objective of the study is written in the Methods instead of the Background?
- The Results section is not clear. Among the 3019 cases and 3333 controls, how many are derived from cancer and stroke cases, respectively?
- What is meant by “significantly positively correlated with cancer risk”? Does it mean an increase or decrease in risk?
-The conclusion is more like describing the results of this study than a conclusion. Please rephrase.

Introduction
- The authors mentioned that several SNPs have been identified in the eNOS gene but failed to justify why they focused on the rs179983 SNP.
- What is meant by “….especially its polymorphism eNOS, as well as…..”?
-What is meant by “…and both cancer risks” in the last sentence of this section?

Methods
- What is meant by “The search period was limited from December 2022 to January 2023”? Were only studies published within this period included in the meta-analysis?
- The authors did not include some essential keywords in the literature search, such as “rs1799983”, “carcinoma”, and “cerebrovascular disease”, which are frequently used in scientific publications.
- Please remove the G vs. T allelic model in the statistical analysis, as it did not yield any meaningful results. The T. vs. G allelic model is sufficient to determine if the recessive T allele is a risk factor for cancer or stroke susceptibility.
- The authors mentioned that a publication bias test cannot be performed due to fewer than 10 studies. This is incorrect, as more than or equal to 3 studies are sufficient for a publication bias test.
- The quality scoring of the studies is not included in the meta-analysis.
- A sensitivity analysis is not included in the meta-analysis.

Results
- It is very confusing to look at the Figure 1 alone. For example, the reports assessed for eligibility are 43, after excluding 21 reports, it should remain at 22. Why are studies included in the meta-analysis only 15?
-The caption for Figures 2 and 3 is not clear. What does A-E mean?
- Many studies published between the years 2012-2023 are not included in the meta-analysis. A few examples are listed below. This shows that the literature search is not comprehensive, probably due to the keywords used.
i) Fadi et al. 2018. World Journal of Neuroscience, 8(1): 98-107.
ii) Phneh et al. 2019. Medicine & Health, 14(1): 118-134.
iii) Jelel et al. 2020. Biological Research for Nursing, 23(3): 408-17.

Discussion
- The second, sixth, and seventh paragraphs are restating the findings of previous studies; they are not comparing to the data of this meta-analysis.
- The last paragraph should be removed and replaced with a paragraph that states the limitations of this study.

Are the rationale for, and objectives of, the Systematic Review clearly stated?

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

Partly
Is the statistical analysis and its interpretation appropriate?

Partly
Are the conclusions drawn adequately supported by the results presented in the review?

Partly

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Gene polymorphisms, meta-analysis, risk association, molecular epidemiology, medical biotechnology

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 3

VERSION 3 PUBLISHED 13 Nov 2023

Open Peer Review

Reviewer Status

Reviewer Reports

	Invited Reviewers
	1	2	3	4
Version 3 (revision) 10 Jun 24			read	read
Version 2 (revision) 28 Mar 24		read
Version 1 13 Nov 23	read	read

Eric Tzyy Jiann Chong, Universiti Malaysia Sabah, Sabah, Malaysia
Katrina Miranda, University of Arizona, Arizona, USA
Mirza Saqib Baig, Indian institute of technology indore, Indore, India
Saori Furuta, University of Toledo Health Science Campus, MetroHealth Medical Center, Case Western Reserve University School of Medicine, Toledo, Cleveland, USA

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

1 Views

31 Oct 2024 | for Version 3

Saori Furuta, Department of Cell & Cancer Biology, College of Medicine and Life Sciences, University of Toledo Health Science Campus, MetroHealth Medical Center, Case Western Reserve University School of Medicine, Toledo, Cleveland, USA

1 Views Cite this report Responses(0)

Not Approved

In the present study, Tualeka et al. explored the association of eNOS 894G>T (Asp298Glu) Polymorphism with elevated risks of cancer and stroke through meta-analysis of published studies. While the pathogenic relevance of this polymorphism to various diseases has been extensively studied, its relevance to stroke and cancer has not been fully determined yet. Their results concluded that there is a significant correlation of this polymorphism to both stoke and cancer, suggesting the roles of this polymorphism as the susceptibility marker for both conditions. Despite the potential significance of the study findings, there are several caveats described below that make it the present form of the manuscript unsuitable for publication. Major revisions would be suggested to improve the quality of the manuscript.
Specific comments:

Please describe the biological consequence of this polymorphism. Several studies showed that this polymorphism reduces the basal level of NO production. If so, the proposed models in Fig. 5 based on the elevated NO production is not correct.
Please describe the nature of studies utilized in tables, including gender, age, disease condition, etc.
Please separate analyses based on cancer types (e.g., tissues and grades).
Please describe all the acronyms in the text, figures, and tables.

Are the rationale for, and objectives of, the Systematic Review clearly stated?

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

No
Is the statistical analysis and its interpretation appropriate?

I cannot comment. A qualified statistician is required.
Are the conclusions drawn adequately supported by the results presented in the review?

Partly
If this is a Living Systematic Review, is the ‘living’ method appropriate and is the search schedule clearly defined and justified? (‘Living Systematic Review’ or a variation of this term should be included in the title.)

No

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Breast cancer, lung cancer, cancer immunology, cancer metabolism, microbiome

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)

Back to all reports

Reviewer Report

3 Views

06 Aug 2024 | for Version 3

Mirza Saqib Baig, Department of Biosciences and Biomedical Engineering, Indian institute of technology indore, Indore, India

3 Views Cite this report Responses(0)

Approved With Reservations

The article entitled "Association of eNOS rs1799983 (894G>T) Polymorphism with Cancer and Stroke Risk: A Meta-Analysis" has derived a meta-analysis study of the effect of different genotypic variants on the function of eNOS and its subsequent role in the risk of Cancer and Stroke. Although the study shows the role of different genotypic variants of eNOS G894T in the outcome of the disease, a few corrections are needed:
1. As most of the genotypes showed heterogeneity in the study, further analysis of the genotypes by deriving different subgroups from the study (e.g., age, sex, ethnicity) can help in identifying the main causes associated with the disease.
2. As the funnel plot showed publication bias in the study, the authors should provide a discussion on the steps to be taken to reduce the publication bias in the study.
3. It is not clear from where the values of the total and the events were given, as the values does not match the individual genotype and allele values in the case and control provided in the Table 2. Author should provide the text in the discussion how the value of the event and total were provided in the table corresponding to each study. Also, how the weight was assigned in the individual study in Figure 2 and 3.
5. The author should provide short discussion on the individual terms such as I², pH, pE etc and why it was used in the Table 2 and 3
6. Author should review the Figure 5 and 6 as the text provided in the discussion doesn’t correlate with the actual figure. Also, it is not clear in Figure 6, whether variant of eNOS leads to more uncoupling of the protein and the disease phenotype compared to control.
7. The author should provide more references of the actual in-vitro studies done on the eNOS G894T variant in the risk of cancer and stroke. Also, there should be more discussion in the text by which mechanism the eNOS G894T variant shows the disease phenotype in cancer and stroke.

Are the rationale for, and objectives of, the Systematic Review clearly stated?

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

Yes
Is the statistical analysis and its interpretation appropriate?

Yes
Are the conclusions drawn adequately supported by the results presented in the review?

No

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, however I have significant reservations, as outlined above.

Respond to this report

Responses (0)

Back to all reports

Reviewer Report

7 Views

02 May 2024 | for Version 2

Katrina Miranda, University of Arizona, Arizona, USA

7 Views Cite this report Responses(0)

Not Approved

The manuscript is highly repetitive, cursory in nature and does not provide any major insights. The authors did not address the concerns of the reviewers, thus I still would recommend rejection.

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

NO and cancer

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)

Back to all reports

Reviewer Report

25 Views

16 Feb 2024 | for Version 1

Katrina Miranda, University of Arizona, Arizona, USA

25 Views Cite this report Responses(0)

Not Approved

The manuscript by Tualeka and colleagues provides a meta-analysis on the correlation of a polymorphism in the eNOS gene on cancer and stroke. The analysis itself is fine. However, presentation of the complex relationship of NO and cancer or stroke is cursory. References include randomly chosen reviews rather than the original literature. It would be beneficial to include an expert on NO in preparation of a revision. For example, the levels of NO produced by the isoforms is not well presented. The polymorphism is also not described sufficiently. Lately, in Figure 5, if eNOs is uncoupled, where does the NO come from to produce peroxynitrite? In sum, more details are needed in this manuscript.

Are the rationale for, and objectives of, the Systematic Review clearly stated?

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

Yes
Is the statistical analysis and its interpretation appropriate?

Yes
Are the conclusions drawn adequately supported by the results presented in the review?

Partly

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

NO and cancer

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)

Back to all reports

Reviewer Report

32 Views

12 Feb 2024 | for Version 1

Eric Tzyy Jiann Chong, Biotechnology Research Institute, Universiti Malaysia Sabah, Sabah, Malaysia

32 Views Cite this report Responses(0)

Not Approved

The manuscript by Tualeka et al. aims to associate the eNOS rs1799983 SNP with the risk of cancer and stroke using a meta-analysis approach. The authors concluded that this SNP is associated with the risk of cancer and stroke in some genetic model comparisons. Overall, the manuscript is poorly written, with many unclear statements and grammatical errors, although it has been reviewed and edited by many authors. There are many concerns that should be addressed by the authors.

Title
- The current title is not specific enough to reflect the contents of the manuscript. Please include the specific gene and polymorphism in the title.

Abstract
-Why is the objective of the study is written in the Methods instead of the Background?
- The Results section is not clear. Among the 3019 cases and 3333 controls, how many are derived from cancer and stroke cases, respectively?
- What is meant by “significantly positively correlated with cancer risk”? Does it mean an increase or decrease in risk?
-The conclusion is more like describing the results of this study than a conclusion. Please rephrase.

Introduction
- The authors mentioned that several SNPs have been identified in the eNOS gene but failed to justify why they focused on the rs179983 SNP.
- What is meant by “….especially its polymorphism eNOS, as well as…..”?
-What is meant by “…and both cancer risks” in the last sentence of this section?

Methods
- What is meant by “The search period was limited from December 2022 to January 2023”? Were only studies published within this period included in the meta-analysis?
- The authors did not include some essential keywords in the literature search, such as “rs1799983”, “carcinoma”, and “cerebrovascular disease”, which are frequently used in scientific publications.
- Please remove the G vs. T allelic model in the statistical analysis, as it did not yield any meaningful results. The T. vs. G allelic model is sufficient to determine if the recessive T allele is a risk factor for cancer or stroke susceptibility.
- The authors mentioned that a publication bias test cannot be performed due to fewer than 10 studies. This is incorrect, as more than or equal to 3 studies are sufficient for a publication bias test.
- The quality scoring of the studies is not included in the meta-analysis.
- A sensitivity analysis is not included in the meta-analysis.

Results
- It is very confusing to look at the Figure 1 alone. For example, the reports assessed for eligibility are 43, after excluding 21 reports, it should remain at 22. Why are studies included in the meta-analysis only 15?
-The caption for Figures 2 and 3 is not clear. What does A-E mean?
- Many studies published between the years 2012-2023 are not included in the meta-analysis. A few examples are listed below. This shows that the literature search is not comprehensive, probably due to the keywords used.
i) Fadi et al. 2018. World Journal of Neuroscience, 8(1): 98-107.
ii) Phneh et al. 2019. Medicine & Health, 14(1): 118-134.
iii) Jelel et al. 2020. Biological Research for Nursing, 23(3): 408-17.

Discussion
- The second, sixth, and seventh paragraphs are restating the findings of previous studies; they are not comparing to the data of this meta-analysis.
- The last paragraph should be removed and replaced with a paragraph that states the limitations of this study.

Are the rationale for, and objectives of, the Systematic Review clearly stated?

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

Partly
Is the statistical analysis and its interpretation appropriate?

Partly
Are the conclusions drawn adequately supported by the results presented in the review?

Partly

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Gene polymorphisms, meta-analysis, risk association, molecular epidemiology, medical biotechnology

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] Abedinzadeh M, Dastgheib SA, Maleki H, et al.: Association of endothelial nitric oxide synthase gene polymorphisms with susceptibility to prostate cancer: A comprehensive systematic review and meta-analysis. Urol. J. 2020; 17(4): 329–337. PubMed Abstract | Publisher Full Text

[2] Adibmanesh A, Bijanzadeh M, Mohammadzadeh G, et al.: The correlation of endothelial nitric oxide synthase gene rs1799983 polymorphisms with colorectal cancer. Int. J. Cancer Manag. 2020; 13(5). Publisher Full Text

[3] Akbar KR, Permatasari AI, Suwarno B, et al.: Lack of association between endothelial nitric oxide synthase (eNOS) G894T gene polymorphism and the risk of bladder cancer Α meta-analysis. Arch. Hell. Med. 2022; 39(5): 647–653.

[4] Akhter MS, Biswas A, Rashid H, et al.: Screening of the NOS3 gene identifies the variants 894G/T, 1998C/G and 2479G/A to be associated with acute onset ischemic stroke in young Asian Indians. J. Neurol. Sci. 2014; 344(1–2): 69–75. PubMed Abstract | Publisher Full Text

[5] Anliaçik SÖ, Tokgöz S, Zamani AG, et al.: Investigation of the relationship between ischemic stroke and endothelial nitric oxide synthase gene polymorphisms [G894T, intron 4 VNTR and T786C]. Turk. J. Med. Sci. 2019; 49(2): 589–594. PubMed Abstract | Publisher Full Text | Free Full Text

[6] Aouf S, Laribi A, Gabbouj S, et al.: Contribution of Nitric oxide synthase 3 genetic variants to nasopharyngeal carcinoma risk and progression in a Tunisian population. Eur. Arch. Otorhinolaryngol. 2019; 276(4): 1231–1239. Publisher Full Text

[7] Branković A, Brajušković G, Nikolić Z, et al.: Endothelial nitric oxide synthase gene polymorphisms and prostate cancer risk in serbian population. Int. J. Exp. Pathol. 2013; 94(6): 355–361. PubMed Abstract | Publisher Full Text | Free Full Text

[8] Buldreghini E, Mahfouz RZ, Vignini A, et al.: Single Nucleotide Polymorphism (SNP) of the Endothelial Nitric Oxide Synthase (eNOS) Gene (Glu298Asp Variant) in Infertile Men With Asthenozoospermia. J. Androl. 2010; 31(5): 482–488. PubMed Abstract | Publisher Full Text

[9] Carkic J, Nikolic N, Nisevic J, et al.: Endothelial nitric oxide synthase polymorphisms/haplotypes are strong modulators of oral cancer risk in serbian population. J. Oral Sci. 2020; 62(3): 322–326. PubMed Abstract | Publisher Full Text

[10] Ben Chaaben A, Mariaselvam C, Salah S, et al.: Polymorphisms in oxidative stress-related genes are associated with nasopharyngeal carcinoma susceptibility. Immunobiology. 2015; 220(1): 20–25. Publisher Full Text

[11] da Costa Escobar Piccoli J , Manfredini V, Hamester FI, et al.: Interaction Between Endothelial Nitric Oxide Synthase Gene Polymorphisms (-786T>C, 894G>T and Intron 4 a/b) and Cardiovascular Risk Factors in Acute Coronary Syndromes. Arch. Med. Res. 2012; 43(3): 205–211. PubMed Abstract | Publisher Full Text

[12] Diakite B, Hamzi K, Slassi I, et al.: G894T endothelial nitric oxide synthase polymorphism and ischemic stroke in Morocco. Meta Gene. 2014; 2(1): 349–357. PubMed Abstract | Publisher Full Text | Free Full Text

[13] Gào X, Schöttker B: Reduction-oxidation pathways involved in cancer development: A systematic review of literature reviews. Oncotarget. 2017; 8(31): 51888–51906. PubMed Abstract | Publisher Full Text | Free Full Text

[14] Gào X, Xuan Y, Benner A, et al.: Nitric Oxide Metabolites and Lung Cancer Incidence: A Matched Case-Control Study Nested in the ESTHER Cohort. Oxidative Med. Cell. Longev. 2019; 2019: 1–9. PubMed Abstract | Publisher Full Text | Free Full Text

[15] El Gohary EA, El Azab AA, Kamal El-Din MMM: Study of Relationship between G894T Variant of Endothelial Nitric Oxide Synthase Gene and Acute Ischemic Stroke. Egyptian J. Hosp. Med. 2017; 69(1): 1607–1613. Publisher Full Text

[16] Guo X: Endothelial nitric oxide (eNOS) gene G894T and VNTR polymorphisms are closely associated with the risk of ischemic stroke development for Asians: Meta-analysis of epidemiological studies. Mol. Biol. Rep. 2014; 41(4): 2571–2583. PubMed Abstract | Publisher Full Text

[17] Hao Y, Montiel R, Huang Y: Endothelial nitric oxide synthase (eNOS) 894 G>T polymorphism is associated with breast cancer risk: A meta-analysis. Breast Cancer Res. Treat. 2010; 124(3): 809–813. PubMed Abstract | Publisher Full Text

[18] Hinz J, Schöndorf D, Bireta C, et al.: The eNOS 894G/T gene polymorphism and its influence on early and long-term mortality after on-pump cardiac surgery. J. Cardiothorac. Surg. 2013; 8(1): 199. PubMed Abstract | Publisher Full Text | Free Full Text

[19] Hung WC, Wu TF, Ng SC, et al.: Involvement of endothelial nitric oxide synthase gene variants in the aggressiveness of uterine cervical cancer. J. Cancer. 2019; 10(12): 2594–2600. PubMed Abstract | Publisher Full Text | Free Full Text

[20] Jang MJ, Jeon YJ, Kim JW, et al.: Association of eNOS polymorphisms (-786T>C, 4a4b, 894G>T) with colorectal cancer susceptibility in the Korean population. Gene. 2013; 512(2): 275–281. PubMed Abstract | Publisher Full Text

[21] Kang MK, Kim OJ, Jeon YJ, et al.: Interplay between polymorphisms in the endothelial nitric oxide synthase (eNOS) gene and metabolic syndrome in determining the risk of ischemic stroke in Koreans. J. Neurol. Sci. 2014; 344(1–2): 55–59. PubMed Abstract | Publisher Full Text

[22] Kaur K, Uppal A, Kaur A: An exonic G894T variant of endothelial nitric oxide synthase gene as a risk factor for ischemic stroke in North Indians. Acta Neurobiol. Exp. 2015; 75(4): 339–350. PubMed Abstract

[23] Koçer C, Benlier N, Balci SO, et al.: The role of endothelial nitric oxide synthase gene polymorphisms in patients with lung cancer. Clin. Respir. J. 2020; 14(10): 948–955. PubMed Abstract | Publisher Full Text

[24] Korde Choudhari S, Chaudhary M, Bagde S, et al.: Nitric oxide and cancer: A review. World J. Surg. Oncol. 2013; 11: 1–11. PubMed Abstract | Publisher Full Text | Free Full Text

[25] Kumar A, Misra S, Kumar P, et al.: Association between Endothelial nitric oxide synthase G894T gene polymorphism and risk of ischemic stroke in North Indian population: A case-control study. Neurol. Res. 2016; 38(7): 575–579. PubMed Abstract | Publisher Full Text

[26] Lee P-C: A Case-Control Study and Meta-Analysis of the Association of eNOS rs1799983 SNP with Stroke Risk. Med. Health. 2019; 14(1): 118–134. Publisher Full Text

[27] Lim KH, Ancrile BB, Kashatus DF, et al.: Tumour maintenance is mediated by eNOS. Nature. 2008; 452(7187): 646–649. PubMed Abstract | Publisher Full Text | Free Full Text

[28] Özçelik AT, Can Demirdöʇen B, Demirkaya Ş, et al.: Importance of NOS3 genetic polymorphisms in the risk of development of ischemic stroke in the Turkish population. Genet. Test. Mol. Biomarkers. 2014; 18(12): 797–803. PubMed Abstract | Publisher Full Text

[29] Rah H, Jeon YJ, Lee WS, et al.: Association of nitric oxide synthase gene polymorphisms (-786T>C, 4a4b, 894G>T) with primary ovarian insufficiency in Korean women. Maturitas. 2013; 74(2): 160–165. PubMed Abstract | Publisher Full Text

[30] Sachdev K: Clinical Pathology and Clinical Bacteriology. J. Clin. Pathol. Bacteriol. 1999; 22(1). Publisher Full Text

[31] Shyu HY, Chen MH, Hsieh YH, et al.: Association of eNOS and Cav-1 gene polymorphisms with susceptibility risk of large artery atherosclerotic stroke. PLoS One. 2017; 12(3): e0174110–e0174112. PubMed Abstract | Publisher Full Text | Free Full Text

[32] Su CW, Chien MH, Lin CW, et al.: Associations of genetic variations of the endothelial nitric oxide synthase gene and environmental carcinogens with oral cancer susceptibility and development. Nitric Oxide. 2018; 79(May): 1–7. PubMed Abstract | Publisher Full Text

[33] Tsay MD, Hsieh MJ, Wang SS, et al.: Impact of endothelial nitric oxide synthase polymorphisms on urothelial cell carcinoma development. Urol. Oncol. 2019; 37(4): 293.e1–293.e9. PubMed Abstract | Publisher Full Text

[34] Tualeka AR, Jalaludin J, Gasana J, et al.: PRISMA Checklist for Article Association between nitric oxide and cancer and stroke risk: A meta-analysis. Zenodo. 2023. Publisher Full Text

[35] Utispan K, Koontongkaew S: High nitric oxide adaptation in isogenic primary and metastatic head and neck cancer cells. Anticancer Res. 2020; 40(5): 2657–2665. Publisher Full Text

[36] Verim L, Toptaş B, Özkan NE, et al.: Possible relation between the NOS3 gene GLU298ASP polymorphism and bladder cancer in Turkey. Asian Pac. J. Cancer Prev. 2013; 14(2): 665–668. PubMed Abstract | Publisher Full Text

[37] Yadav SK, Gupta S, Yadav A, et al.: Endothelial nitric oxide synthase gene polymorphisms modulate the risk of squamous cell carcinoma of head and neck in north Indian population. Meta Gene. 2019; 21(October 2018): 100575. Publisher Full Text

[38] Yanar K, Çakatay U, Aydin S, et al.: Relation between Endothelial Nitric Oxide Synthase Genotypes and Oxidative Stress Markers in Larynx Cancer. Oxidative Med. Cell. Longev. 2016; 2016: 1–8. PubMed Abstract | Publisher Full Text | Free Full Text

[39] Yang C, Hawkins KE, Doré S, et al.: Neuroinflammatory mechanisms of blood-brain barrier damage in ischemic stroke.2019.

[40] Yao YS, Chang WW, Jin YL, et al.: An updated meta-analysis of endothelial nitric oxide synthase gene: Three well-characterized polymorphisms with ischemic stroke. Gene. 2013; 528(2): 84–92. PubMed Abstract | Publisher Full Text

[41] Zhao C, Yan W, Zu X, et al.: Association between endothelial nitric oxide synthase 894G>T polymorphism and prostate cancer risk: a meta-analysis of literature studies. Tumor Biol. 2014; 35(12): 11727–11733. PubMed Abstract | Publisher Full Text

Association between eNOS rs1799983 (894G>T) Polymorphism with Cancer and Stroke Risk: A meta-analysis

Abstract

Background

Methods

Results

Conclusions

Keywords

Revised Amendments from Version 2

Introduction

Methods

Search strategy

Inclusion and exclusion standard

Data extraction

Statistical analysis

Results

Study processing

Figure 1. Flow diagram depicting the inclusion process of the studies in the meta-analysis.

Table 1. Characteristics and genetic frequencies derived the studies included in the meta-analysis.

Meta-analysis

Table 2. The relationship of cancer risk with eNOS G894T polymorphism in summary.

Table 3. The relationship of stroke risk with eNOS G894T polymorphism in summary.

Heterogeneity among studies

Figure 2. Forest plots depicting the association of eNOS 894G>T polymorphisms with cancer risk in all genetic models, including: A). T vs G, B). GT + TT vs GG, C). TT vs GG + GT, and D). GT vs GG + TT, respectively.

Figure 3. Forest plots illustrating the association of eNOS 894G>T polymorphisms with stroke risk in all genetic models, including: A). T vs G, B). GT + TT vs GG, C). TT vs GG + GT, and D). GT vs GG + TT, respectively.

Publication bias

Figure 4. Funnel Plot of association between eNOS 894G>T polymorphism and risk of cancer and stroke.

Discussion

Figure 5. The role of nitric oxide in cancer.

Figure 6. The role of nitric oxide in stroke.

Conclusions

Data availability

Underlying data

Reporting guidelines

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