Lin SY, Huang H, Yu JJ, Su F, Jiang T, Zhang SY, Lv L, Long T, Pan HW, Qi JQ, Zhou Q, Tang WF, Ding GW, Wang LM, Tan LJ, Yin J. Activin A receptor type 1C single nucleotide polymorphisms associated with esophageal squamous cell carcinoma risk in Chinese population. World J Gastrointest Oncol 2025; 17(1): 96702 [DOI: 10.4251/wjgo.v17.i1.96702]
Corresponding Author of This Article
Jun Yin, MD, PhD, Professor, Department of Thoracic Surgery, Zhongshan Hospital Affiliated to Fudan University, No. 180 Fenglin Road, Xuhui District, Shanghai 200032, China. jun_yin@fudan.edu.cn
Research Domain of This Article
Genetics & Heredity
Article-Type of This Article
Case Control Study
Open-Access Policy of This Article
This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Si-Yun Lin, Jin-Jie Yu, Feng Su, Tian Jiang, Shao-Yuan Zhang, Li-Jie Tan, Jun Yin, Department of Thoracic Surgery, Zhongshan Hospital Affiliated to Fudan University, Shanghai 200032, China
Si-Yun Lin, Hou Huang, Shanghai Key Laboratory of Clinical Geriatric Medicine, Huadong Hospital Affiliated to Fudan University, Shanghai 200040, China
Lu Lv, Tao Long, Hui-Wen Pan, Jun-Qing Qi, Guo-Wen Ding, Department of Cardiothoracic Surgery, The Affiliated People's Hospital of Jiangsu University, Zhenjiang 212002, Jiangsu Province, China
Qiang Zhou, Department of Thoracic Surgery, Sichuan Cancer Hospital & Institute, Chengdu 610042, Sichuan Province, China
Wei-Feng Tang, Department of Cardiothoracic Surgery, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing 210000, Jiangsu Province, China
Li-Ming Wang, Department of Respiratory and Critical Care Medicine, Shanghai Xuhui Central Hospital, Shanghai 200032, China
Author contributions: Yin J and Tan LJ conceptualized and designed the research; Yin J, Lv L, Long T, Pan HW, Qi JQ, Zhou Q and Tang WF screened patients and acquired clinical data; Yin J, Lv L and Long T collected blood specimen and performed laboratory analysis; Huang H and Lin SY performed Data analysis and wrote the first draft of the manuscript; Lin SY was responsible for data re-analysis and re-interpretation, figure plotting, comprehensive literature search, preparation and submission of the current version of the manuscript; Yu JJ, Su F and Zhang SY provided suggestions on the methodology and writing guidance; Jiang T and Wang LM helped revise the manuscript; Ding GW, Tan LJ and Yin J were responsible for study administration; All the authors have read and approved the final manuscript. Both Lin SY and Huang H have made crucial and indispensable contributions towards the completion of the project and thus qualified as the co-first authors of the paper. Both Tan LJ and Yin J have played important and indispensable roles in the experimental design, data interpretation and manuscript preparation as the co-corresponding authors. Yin J and Wang LM applied for and obtained the funds for this research project.
Supported byThe National Natural Science Foundation of China, No. 82350127 and No. 82241013; the Shanghai Natural Science Foundation, No. 20ZR1411600; the Shanghai Shenkang Hospital Development Center, No. SHDC2020CR4039; the Bethune Ethicon Excellent Surgery Foundation, No. CESS2021TC04; and Xuhui District Medical Research Project of Shanghai, No. SHXH201805.
Institutional review board statement: The study was reviewed and approved by the Ethics Committee of Jiangsu University (Zhenjiang, China) and the Review Board of the institution (Approval No. K20160036-W).
Informed consent statement: All recruited participants were provided with informed consent.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: The dataset generated and analyzed during the current study are available in 1000Genomes project [1000G: PRJEB6930] from the NCBI SNP database. The genotype data of the selected SNPs used to support the findings of this study are available from the corresponding author on reasonable request at jun_yin@fudan.edu.cn.
STROBE statement: The authors have read the STROBE Statement—checklist of items, and the manuscript was prepared and revised according to the STROBE Statement—checklist of items.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Jun Yin, MD, PhD, Professor, Department of Thoracic Surgery, Zhongshan Hospital Affiliated to Fudan University, No. 180 Fenglin Road, Xuhui District, Shanghai 200032, China. jun_yin@fudan.edu.cn
Received: May 13, 2024 Revised: September 1, 2024 Accepted: October 14, 2024 Published online: January 15, 2025 Processing time: 212 Days and 20.3 Hours
Abstract
BACKGROUND
Transforming growth factor-β (TGF-β) superfamily plays an important role in tumor progression and metastasis. Activin A receptor type 1C (ACVR1C) is a TGF-β type I receptor that is involved in tumorigenesis through binding to different ligands.
AIM
To evaluate the correlation between single nucleotide polymorphisms (SNPs) of ACVR1C and susceptibility to esophageal squamous cell carcinoma (ESCC) in Chinese Han population.
METHODS
In this hospital-based cohort study, 1043 ESCC patients and 1143 healthy controls were enrolled. Five SNPs (rs4664229, rs4556933, rs77886248, rs77263459, rs6734630) of ACVR1C were assessed by the ligation detection reaction method. Hardy-Weinberg equilibrium test, genetic model analysis, stratified analysis, linkage disequilibrium test, and haplotype analysis were conducted.
RESULTS
Participants carrying ACVR1C rs4556933 GA mutant had significantly decreased risk of ESCC, and those with rs77886248 TA mutant were related with higher risk, especially in older male smokers. In the haplotype analysis, ACVR1C Trs4664229Ars4556933Trs77886248Crs77263459Ars6734630 increased risk of ESCC, while Trs4664229Grs4556933Trs77886248Crs77263459Ars6734630 was associated with lower susceptibility to ESCC.
CONCLUSION
ACVR1C rs4556933 and rs77886248 SNPs were associated with the susceptibility to ESCC, which could provide a potential target for early diagnosis and treatment of ESCC in Chinese Han population.
Core Tip: In this study, a case-control approach was used to investigate the association between specific single nucleotide polymorphisms (SNPs) of the ACVR1C gene and the risk of esophageal squamous cell carcinoma (ESCC) in a Chinese Han population. The study included 2186 participants, comprising 1043 ESCC patients and 1143 healthy controls. Notably, the rs4556933 G>A SNP was significantly associated with ESCC risk across several genetic models. Stratified analysis indicated an increased risk in older males and smokers. These findings suggest that ACVR1C SNPs play a critical role in ESCC susceptibility, meriting further investigation into their involvement in cancer development.
Citation: Lin SY, Huang H, Yu JJ, Su F, Jiang T, Zhang SY, Lv L, Long T, Pan HW, Qi JQ, Zhou Q, Tang WF, Ding GW, Wang LM, Tan LJ, Yin J. Activin A receptor type 1C single nucleotide polymorphisms associated with esophageal squamous cell carcinoma risk in Chinese population. World J Gastrointest Oncol 2025; 17(1): 96702
Esophageal cancer (EC) is a common malignancy of the upper digestive tract. The International Agency for Research on Cancer reported approximately 604000 new cases of EC, ranking seventh, and 540000 deaths, ranking sixth, worldwide in 2020[1]. The incidence of EC varies regionally, with China presenting a high-incidence area, with new cases accounting for over half of the global total count[2]. Moreover, in contrast to patients of Western origin, the pathological type detected in most Chinese patients is esophageal squamous cell carcinoma (ESCC).
The onset of ESCC is frequently inconsistent. Genetic, epigenetic, and environmental factors significantly contribute to ESCC pathogenesis; smoking and alcohol consumption are crucial exogenous risk factors[3]. Additionally, unhealthy dietary habits, such as hot eating, uneven intake, and excessive intake of pickled products, can increase the risk of tumorigenesis. Gene and chromosomal abnormalities are closely related to the occurrence and development of heterogeneity, a major characteristic of tumors. Therefore, further genetic studies are necessary to explore the pathogenic processes of ESCC and its association with genetic abnormalities, including single nucleotide polymorphisms (SNP), the most common type of genetic variation.
Activin A receptor type 1C (ACVR1C) is a transforming growth factor-β (TGF-β) type I receptor, with a single-span transmembrane domain, which is also known as activin receptor-like kinase 7 (ALK7). The receptors for TGF-β superfamily ligands comprise quadruplex complexes with two homodimers of type I and type II subunits[4]. More than thirty-five TGF-β superfamily receptors have been found in mammals[5,6]. The TGF-β superfamily includes TGF-β1-3, activins A and B, inhibins, growth differentiation factors, bone morphogenetic proteins, nodal proteins, and other ligand components; they are involved in cell proliferation, differentiation, adhesion, wound healing, and immune responses through binding in various ways[7]. Numerous studies have demonstrated the magnitude of TGF-β superfamily cytokines during tumor progression; the differential activities of TGF-βs and their core receptors in metastatic phenotypes present the most crucial part of its involvement in tumorigenesis[8]. Therefore, the regular expression and functions of the TGF-β superfamily play a critical role in maintaining the stability of the organism.
ACVR1C is the seventh and most recently identified member of the type-I receptor subfamily. ACVR1C is highly expressed in tissues and cell types with endocrine or neuroendocrine functions compared with other TGF-β receptors, thus exhibiting its correlation with obesity, type II diabetes, and metabolic syndrome[9]. Early studies elucidated the large amount of mRNA expression encoding ALK7 in the adipose tissue, gastrointestinal tract, ovary, prostate, pancreas, and a few brain nuclei, especially in the cerebellum[10-12]. Moreover, it is involved in cancer-related apoptosis and cell proliferation in vitro. ACVR1C and its ligand Nodal have pro-apoptotic effects in various cancer cells, including breast cancer[13,14]. However, contrasting reports are also available[15,16]. Additionally, ACVR1C signaling acts as a homeostatic tissue barrier against tumorigenesis and metastasis[17]. Consequently, further research can clarify the pro-and anti-tumorigenic effects of ACVR1C.
Hence, in this study, we hypothesized that some SNPs of ACVR1C might be correlated with the susceptibility to ESCC and aimed to determine their correlation with esophageal tumorigenesis using an association analysis of candidate SNPs. We aimed to verify this hypothesis through a hospital-based cohort study in a Chinese Han population.
MATERIALS AND METHODS
Study design
This single-center, case-control study was conducted from October 1, 2008 to January 31, 2017. The Ethics Committee of Jiangsu University (Zhenjiang, China) and the Institutional Review Board (approval No. K20160036-W) approved this study. The research protocol was formulated and executed following the Ethical Principles for Medical Research Involving Human Subjects by the World Medical Association Declaration of Helsinki. All the participants provided informed consent.
A total of 2186 participants from the People's Hospital affiliated with Jiangsu University were retrospectively enrolled. The case group included 1043 patients with ESCC. All diagnoses were confirmed via pathological biopsies. Patients, who underwent neoadjuvant therapy, had other EC histological types, or had a history of other cancers, were excluded from the study. The control group included non-tumor patients with healthy physical examination and trauma during the same period. Propensity matching was conducted considering the distribution characteristics of the case groups such as sex, age, and residential area.
Demographic characteristics, including sex, age, and ESCC-related risk factors (smoking status and alcohol consumption), were recorded based on a questionnaire. Smoking was defined as the active smoking of ≥ 1 cigarette per day for one year. Alcohol consumption was defined as drinking at least three times per week for more of over six months. After obtaining the informed consent, 2 mL of fasting venous blood was drawn from each participant for subsequent tests.
DNA extraction and genotyping
The genomic DNA was extracted from peripheral blood using the QIAamp DNA Blood Mini Kit (Qiagen, Berlin, Germany). The ligation detection reaction (LDR) method was used for further SNP genotyping analysis with technical support from Genesky Biotechnology Inc. (Shanghai, China). Meanwhile, the consistency of all data was confirmed using 10% of the samples that were randomly selected. Double-track data entries were used to ensure the authenticity. To select the correlated SNPs loci and tagSNPs for further analyses, a pilot linkage disequilibrium (LD) analysis was performed using data sourced from the 1000Genomes project (1000G: PRJEB6930) of the National Center for Biotechnology Information SNP database.
Statistical analysis
SPSS 24.0 (Chicago, IL, United States), Haploview4.1, SHEsis software (online version)[18] were used to analyze the data. Hardy-Weinberg equilibrium (HWE) test was conducted to analyze the genotype distribution frequency of the control group by χ2 test. SNPs in the control group with a P value greater than 0.05 were considered to be consistent with HWE, and could be further analyzed. χ2 test was also applied to compare the differences in demographic characteristics between the ESCC case and control group.
Diverse genetic models and subgroup-stratified analyses were performed using logistic regression. Assuming alleles A and B are located in a single SNP: (1) Co-dominant model (risk associated with AB individuals lies between that of AA and BB individuals); (2) Dominant model (risk increased by allele B); (3) Recessive model (risk increased by two copies of allele B); and (4) Additive model (risk increased by r-fold for AB and 2r for BB) were considered[19]. Odds ratios (OR) and 95%CI were calculated to assess the strength of the association between SNPs and the risk of disease. Subsequently, adjusted OR and corresponding CI were calculated based on hierarchical variables, including age, sex, smoking, and alcohol consumption status. Finally, haplotype and LD analyses were performed using the SHEsis software. The coefficient of LD was evaluated using the D' and r2 values. Bilateral test data were considered statistically significant at P < 0.05.
RESULTS
Characteristics of ESCC patients and healthy controls
The demographic characteristics and ESCC-related risk factors of the enrolled participants are listed in Table 1. We analyzed 1043 patients with ESCC (63.07 ± 7.27 years, mean age ± SD; 758 males and 285 females) and 1143 healthy controls (62.64 ± 9.90 years, mean age ± SD; 828 males and 315 females). Age and sex did not significantly differ between groups. However, smoking and alcohol consumption status significantly differed between the groups (P < 0.001), with higher rates for both parameters in the case group than in the control group.
Table 1 distribution of selected demographic variables and risk factors in esophageal squamous cell carcinoma case and control groups, n (%).
Variable
Case group (n = 1043)
Control group (n = 1143)
P value
Age (years), mean ± SD
63.07 ± 7.27
62.64 ± 9.90
0.252
Age (years)
0.041
≥ 63
572 (54.84)
577 (50.48)
< 63
471 (45.16)
566 (49.52)
Sex
0.903
Male
758 (72.67)
828 (72.44)
Female
285 (27.33)
315 (27.56)
Smoking status
< 0.001
Never
589 (56.47)
810 (70.87)
Ever
454 (43.53)
333 (29.13)
Alcohol consumption
< 0.001
Never
714 (68.46)
1061 (92.82)
Ever
329 (31.54)
82 (7.18)
The basic information on the five selected genotyped SNPs is presented in Table 2. All SNPs were located on chromosome 2. The minor allele frequency (MAF) of SNPs in the control group corresponded to that in the East Asian population, which was reflected in the 1000 Genomes database. The genotyping value was > 95%, indicating the reliability of the experimental results. The HWE test was conducted for the control group, which validated that the study population was in line with the Hardy-Weinberg balance (P > 0.05), and that the study population was representative.
Table 2 Primary information of five selected genotyped single nucleotide polymorphisms of Activin A receptor type 1C: rs4664229 T>C, rs4556933 G>A, rs77886248 T>A, rs77263459 T>C, rs6734630 A>G.
Risk of ACVR1C SNPs with ESCC analyzed by genetic model
The associations between ESCC and each SNP are summarized in Table 3 and Figure 1. Based on four genetic models, the correlation between ESCC and various genotypes was analyzed. rs4556933 G>A was significantly associated with ESCC in the dominant, recessive, and additive models (P < 0.001, = 0.036, and 0.015, respectively). In addition, in the co-dominant model, the GA genotype of rs4556933 was associated with a lower risk of ESCC. The significant association between rs77886248 T>A and ESCC was reflected in only the dominant model (P = 0.038), which suggests that the TA and AA genotypes significantly increased the risk of ESCC between the case and healthy control groups. Nevertheless, genetic models showed no association of ESCC with rs4664229, rs77263459, and rs6734630 were not associated with ESCC, which is attributable to the marginal statistical significance observed in the genotype frequencies. The genotype distributions of the two significant SNPs are shown in Figure 2.
Figure 1 Forest plots of genetic modeling to analyze the risk of activin A receptor type 1C single nucleotide polymorphisms with esophageal squamous cell carcinoma.
OR: Odds ratio.
Figure 2 Genotype distributions of two significant single nucleotide polymorphisms.
A: Rs4556933; B: Rs77886248.
Table 3 Analyses of associations between selected genotyped single nucleotide polymorphisms of activin A receptor type 1C on risk of esophageal squamous cell carcinoma.
Locus
Genotype
Control
Case
Co-dominant model
Dominant model
Recessive model
Addictive model
OR (95%CI)
P value
OR (95%CI)/P value
OR (95%CI)/P value
OR (95%CI)/P value
rs4664229
TT
908
835
1
0.311
0.894 (0.721-1.107)
1.487 (0.624-3.545)
0.928 (0.76-1.131)
TC
221
177
0.871 (0.7-1.084)
0.216
0.303
0.37
0.456
CC
9
12
1.45 (0.608-3.459)
0.402
rs4556933
GG
550
587
1
< 0.001
0.684 (0.577-0.811)
1.349 (1.019-1.786)
0.848 (0.743-0.969)
GA
487
311
0.598 (0.498-0.719)
< 0.001
< 0.001
0.036
0.015
AA
101
118
1.095 (0.819-1.463)
0.541
rs77886248
TT
1066
936
1
0.087
1.408 (1.019-1.944)
0.555 (0.05-6.126)
1.366 (0.999-1.870)
TA
70
88
1.432 (1.034-1.983)
0.031
0.038
0.631
0.051
AA
2
1
0.569 (0.052-6.29)
0.646
rs77263459
TT
647
593
1
0.087
0.958 (0.807-1.136)
1.169 (0.84-1.628)
0.999 (0.870-1.147)
TC
417
354
0.926 (0.773-1.109)
0.405
0.62
0.355
0.988
CC
74
77
1.135 (0.810-1.592)
0.462
rs6734630
AA
908
838
1
0.402
0.881 (0.711-1.092)
1.236 (0.5-3.054)
0.906 (0.743-1.105)
GA
221
177
0.868 (0.697-1.08)
0.204
0.247
0.646
0.331
GG
9
10
1.204 (0.487-2.977)
0.688
Stratified analysis of SNPs and the risk of ESCC in different subgroups
The stratified analyses based on sex, age, smoking status, and alcohol consumption better reflected the correlation between ACVR1C SNPs and susceptibility to ESCC (Table 4 and Table 5; Figure 3). In male participants older than 63 years, rs4556933 G>A was statistically significant in the dominant, recessive, and co-dominant models when comparing GG with GA (P < 0.05). Similar results were found in the dominant and co-dominant (TA vs TT) models of rs77886248 T>A (P = 0.027/0.025, with adjusted OR = 1.619/1.623, respectively), indicating that male sex is a risk factor for ESCC. For rs77886248, smoking was also associated with a higher risk of ESCC in the dominant and co-dominant (TA vs TT) models (P = 0.035/0.024, adjusted OR = 1.954/2.401).
Figure 3 Forest plots.
A: Forest plots of stratified analysis to rs4556933 genotype and esophageal squamous cell carcinoma (ESCC) risk; B: Forest plots of stratified analysis to rs77886248 genotype and ESCC risk. OR: Odds ratio.
Table 4 Stratified analyses between rs4556933 polymorphism and esophageal squamous cell carcinoma risk by sex, age, smoking status and alcohol consumption.
Variables
Control/case numbers
Adjusted OR/P value (95%CI of OR)
GG
GA
AA
GA + AA
GG
GA
AA
GA + AA
AA vs (GG + GA)
Sex
Male
399/424
351/225
74/91
425/316
1
0.587/< 0.01
1.18/0.369
0.689/0.001
1.463/0.032
(0.464-0.742)
(0.823-1.691)
(0.555-0.855)
(1.033-2.071)
Female
151/163
136/86
27/27
163/113
1
0.57/0.02
0.898/0.718
0.624/0.005
1.156/0.614
(0.401-0.811)
(0.502-1.607)
(0.449-0.868)
(0.658-2.032)
Age
≥ 63
290/324
241/163
45/66
286/229
1
0.588/< 0.01
1.283/0.257
0.697/0.004
1.582/0.032
(0.449-0.769)
(0.833-1.976)
(0.543-0.893)
(1.041-2.403)
< 63
260/263
246/148
56/52
302/200
1
0.568/< 0.01
0.932/0.754
0.634/0.001
1.198/0.406
(0.427-0.755)
(0.601-1.447)
(0.487-0.826)
(0.783-1.832)
Smoking status
Ever
151/262
152/135
29/52
181/187
1
0.485/< 0.01
1.009/0.974
0.568/0.001
1.39/0.223
(0.344-0.683)
(0.583-1.749)
(0.412-0.782)
(0.818-2.362)
Never
399/325
335/176
72/66
407/242
1
0.645/< 0.01
1.156/0.442
0.734/0.006
1.38/0.078
(0.509-0.817)
(0.799-1.674)
(0.59-0.914)
(0.964-1.973)
Alcohol consumption
Ever
31/190
45/92
6/38
51/130
1
0.326/< 0.01
1.019/0.969
0.407/< 0.01
1.745/0.23
(0.192-0.552)
(0.395-2.63)
(0.246-0.674)
(0.703-4.332)
Never
519/397
442/219
95/80
537/299
1
0.646/< 0.01
1.117/0.507
0.729/0.001
1.343/0.068
(0.524-0.797)
(0.806-1.548)
(0.601-0.885)
(0.978-1.842)
Table 5 Stratified analyses between rs77886248 polymorphism and esophageal squamous cell carcinoma risk by sex, age, smoking status and alcohol consumption.
Variables
Control/case numbers
Adjusted OR/P value (95%CI of OR)
TT
TA
AA
TA + AA
TT
TA
AA
TA + AA
AA vs (TA + AA)
Sex
Male
779/680
44/64
1/1
45/65
1
1.619/0.027
1.772/0.686
1.623/0.025
1.714/0.704
(1.057-2.482)
(0.11-28.458)
(1.063-2.476)
(0.107-27.524)
Female
287/256
26/24
1/0
27/24
1
1.024/0.937
0.979/0.942
(0.57-1.839)
-/- (-)
(0.548-1.749)
-/- (-)
Age
≥ 63
544/509
31/49
1/1
32/50
1
1.643/0.046
1.255/0.874
1.631/0.046
1.21/0.894
(1.009-2.675)
(0.76-20.757)
(1.009-2.639)
(0.073-20.012)
< 63
522/427
39/39
1/0
40/39
1
1.158/0.559
1.134/0.616
(0.707-1.898)
-/- (-)
(0.694-1.853)
-/- (-)
Smoking status
Ever
314/405
18/45
0/1
18/46
1
1.954/0.035
-/- (-)
2.401/0.024
-/- (-)
(1.049-3.642)
(1.099-3.791)
Never
752/531
52/43
2/0
54/0
1
1.164/0.483
-/- (-)
1.118/0.604
-/- (-)
(0.762-1.778)
(0.734-1.702)
Alcohol consumption
Ever
76/294
6/28
0/0
6/28
1
1.111/0.824
-/- (-)
1.111/0.824
-/- (-)
(0.44-2.807)
(0.44-2.807)
Never
990/642
64/60
2/1
66/61
1
1.41/0.068
0.722/0.793
1.389/0.078
0.704/0.777
(0.975-2.038)
(0.64-8.61)
(0.964-1.999)
(0.062-7.959)
Haplotype analysis of SNPs and susceptibility to ESCC
As listed in Table 6 and Table 7, the haplotype analysis of the five SNPs showed that Trs4664229Grs4556933Trs77886248Trs77263459Ars6734630 was the most common genotype in all participants, included in the case and control groups (58.7% and 57.4%, respectively). Trs4664229Ars4556933Trs77886248Crs77263459Ars6734630 was considered to increase the risk of ESCC (P = 0.012, OR = 1.211), whereas, Trs4664229Grs4556933Trs77886248Crs77263459Ars6734630 was associated with lower susceptibility to ESCC (P < 0.01, OR = 0.126).
Table 6 Haplotype frequencies in the case and control group, and risk of esophageal squamous cell carcinoma.
Table 7 Linkage disequilibrium analysis using parameter D and r2.
D’/ r2
rs4556933
rs77886248
rs77263459
rs6734630
rs4664229
0.156/0.001
0.123/0.005
0.333/0.004
0.997/0.985
rs4556933
-
0.993/0.016
0.849/0.588
0.154/0.001
rs77886248
-
-
0.988/0.013
0.125/0.005
rs77263459
-
-
-
0.334/0.004
DISCUSSION
In this study, we demonstrate the relationship between functional ACVR1C polymorphisms and ESCC in a Chinese Han population through an association analysis of candidate SNPs. After collecting blood samples from 2186 participants and conducting LDR analysis, we identified five target SNP loci of AVCR1C. The LD analysis revealed a significant correlation among the five loci, especially between rs4556933 G>A and rs77263459 T>C. These loci are not in the same functional region; rs4556933 is a synonymous variant of the coding sequence and rs77263459 is an intron-associated variant. However, a strong LD is detected (D = 0.849, r2 = 0.588), which indicates a similar influence on gene coding. In this study, the MAF of ACVR1C rs77886248, rs77263459, and rs6734630 in the control group were A = 0.0325, C = 0.2482, and G = 0.1050, respectively, which followed the East Asian population frequency distribution in the 1000Genomes database (A = 0.0387, C = 0.2361, G = 0.0913). The frequency of the first two alleles was significantly higher than that of the global data (A = 0.0078, C = 0.1042), whereas, the third allele showed lower frequency than the global data (G = 0.1865). This demonstrated differences in the frequency distribution of the gene polymorphism loci across species, which may have led to regional diversity in the incidence rate and pathological types of ESCC.
As a receptor binds to the TGF-β superfamily (ligands including Nodal, Activin A/B, and GDF3), ACVR1C is involved in the regulation of tumor progression and metastasis. Asnaghi et al[15] reported that ACVR1C/SMAD2 signaling promotes the invasion and growth of retinoblastoma[15]. However, Lonardo et al[20] suggested that downregulated ACVR1C is a marker of poor prognosis[20]. A significant association of ACVR1C rs4556933 G>A and rs77886248 T>A with ESCC was observed in certain genotypes and genetic models. In the co-dominant test, the GA genotype of rs4556933 reduced the risk of ESCC compared to the wild-type GG genotype (P < 0.001, OR = 0.598, 95%CI: 0.498-0.719); however, the TA genotype of rs77886248 served an independent risk factor for ESCC (P < 0.001, OR = 1.432, 95%CI: 1.034-1.983). However, compared to the wild-type genotype in the co-dominant model, the homozygous mutations of the selected SNPs exhibited no significant association with the risk of ESCC, which is potentially attributed to the hereditary modes and complex mechanisms of SNP in tumor development and progression. Moreover, rs4556933 G>A significantly increased the risk of ESCC in the recessive test, suggesting that the effect of the homozygous mutant AA differs from that of the GA genotype. The hazardous effects of homozygous mutations are associated with tumor immune escape[21]. SNP polymorphism usually involves the transition or inversion-induced variation at a single base. Hence, haplotype analysis further validated the relationship between the five loci and ESCC risk.
Stratified analyses of the SNP rs4556933 revealed a significantly reduced risk of ESCC in subjects with a GA mutation, regardless of their subgroup. For males (age > 63 years) with AA mutants, the risk of ESCC increased, which is consistent with an epidemiological report in China[22]. Only a few studies have reflected the clinical significance of rs4556933 G>A; its correlation with preeclampsia pathogenesis was first reported in a Norwegian population[23]. The genetic function of rs4556933 reflects that it is a synonymous variant; it is considered a silent mutation owing to no alterations in protein sequences. As synonymous mutations are well accepted to be neutral or nearly neutral, they are often neglected in research on pathogenic or protective mutations. However, synonymous mutations in the oncogene KRAS were reported to have a relevant impact on gene expression and mRNA secondary structure[24]. Furthermore, synonymous variants can affect gene regulation via transcription, splicing, mRNA stability, and translation, inducing loss of function and poor prognosis in renal cell carcinoma[25,26]. Therefore, the correlation between this mutation and tumorigenesis may have been underestimated. Additionally, an approximately two-fold increase in the risk of ESCC was detected in older smokers with the mutant genotype (TA and AA) of rs77886248 than in those with wild-type TT, which was consistent with a previous report[27].
Currently, neoadjuvant chemoradiotherapy or chemotherapy combined with surgical resection is considered the standard treatment for patients with locally advanced EC[28]. With the progress in immunotherapy, immune checkpoint inhibitors have been applied in the adjuvant and preoperative treatment stages, ranging from second-line treatment to advanced stages, all of which enhanced the pathologic complete response rate and prolonged disease-free survival[29,30]. Therefore, identifying targets for efficacy assessments may help screen the potential benefit populations and provide tailored therapeutic strategies for patients. Minari et al[31] extracted and analyzed the correlation of SNPs in the programmed death ligand-1 (PD-L1) gene with immunotherapy efficacy in a cohort of 166 non-small cell lung cancer cases, and demonstrated an association of the PD-L1 rs4143815 SNP with a long clinical benefit cohort (P = 0.02)[31]. Additionally, SNPs and treatment safety have a potential functional correlation, with 12 mutations or SNPs associated with an increased risk of melanoma immune-related adverse events[32].
Despite advances in novel therapies, the prognosis of EC remains poor. The early symptoms of EC are insidious; most cases exhibit are locally advanced stage when diagnosed, with lymph node metastasis, resulting in a high recurrence rate and poor 5-year survival[33]. Therefore, the timely diagnosis and identification of new therapeutic targets are required. Among all subgroups analyzed in this study, subjects carrying the ACVR1C rs4556933 GA genotype exhibited a significantly reduced risk of ESCC, which can be considered a candidate locus involved in tumorigenesis and provide potential targets for diagnosis and treatment.
Our study has several limitations. First, this was a hospital-based cohort study and the control group might not accurately represent the general population. Second, owing to technical constraints, we could not verify gene expression through functional experiments in cells or animals. Moreover, beyond alcohol and smoking, other risk factors of ESCC, which include diet and nutrition, infection and microbiome, gastric atrophy, and family income, should be considered when defining subgroups. In a large-scale case-control study, the association of SNPs with disease progression will be further explored depending on the availability of complementary treatment modalities, efficacy evaluations, and long-term follow-up data.
CONCLUSION
In summary, the mutant of GA genotype in ACVR1C rs4556933 G>A showed a suggestive association with a lower risk of ESCC, while rs77886248 T>A increased the susceptibility to ESCC, especially in older male smokers of Chinese Han population.
Footnotes
Provenance and peer review: Unsolicited article; Externally peer reviewed.
Peer-review model: Single blind
Specialty type: Oncology
Country of origin: China
Peer-review report’s classification
Scientific Quality: Grade C, Grade C
Novelty: Grade B, Grade C
Creativity or Innovation: Grade B, Grade C
Scientific Significance: Grade B, Grade C
P-Reviewer: Luo X; Zhan X S-Editor: Li L L-Editor: A P-Editor: Zhao YQ
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