Research On The Correlation Between Polymorphisms Of RIG-Ⅰ And ERCC2 Genes With HBV-related Hepatocellular Carcinoma

BackgroundChronic hepatitis B virus(HBV)infection is a major risk factor for the occurrence of hepatocellular carcinoma(HCC)in China,and host genetic susceptibility plays an important role in the occurrence and development of HBV-related HCC(HBV-HCC).Retinoic acid‐inducible gene 1(RIG-I)is a kind of RIG-I-like pattern recognition receptor molecule that activates downstream RIG-I signaling pathways by binding and recognizing virus-derived RNA,stimulating the expression of inflammatory factors and type I interferons,thereby initiating an antiviral innate immune response.Excision repair cross-complementation group 2(ERCC2)is one of the most important members of the nucleotide resection repair(NER)system,which is essential for maintaining genomic stability and is closely related to cancer development.Although some studies have shown that RIG-I and ERCC2 may be closely related to the development of HBV infection,little is known at home and abroad about the correlation between RIG-I and ERCC2 gene polymorphisms and HBV chronic infection and HBV-HCC.ObjectivesTo explore the genetic susceptibility between RIG-I(rs3824456,rs10813831,rs3739674,and rs2777729)gene polymorphisms and ERCC2(rs238406,rs238407,rs238408 and rs1799785)gene polymorphisms and chronic HBV infection and HBVHCC in the Chinese Han population in the Jingzhou area,Hubei Province,and to provide a reference for finding the genetic susceptibility markers of HBV-HCC and chronic HBV infection.MethodsA case-control study was conducted in the Jingzhou area,Hubei Province,with the Chinese Han population as the research object.There were 154 healthy control cases and 325 patients with chronic HBV infection,including 142 patients with HBV-HCC,150 patients with chronic hepatitis B(CHB),and 33 patients with hepatitis B-related liver cirrhosis(LC).Human DNA was extracted from whole blood and sequenced after polymerase chain reaction(PCR)amplification,and the genotypes of RIG-I rs3824456,rs10813831,rs3739674,rs2777729 and ERCC2 rs238406,rs238407,rs238408,rs1799785 genotypes were detected by Sanger sequencing.SPSS26.0 and the SNPStats web tool(https://www.snpstats.net/start.htm)were used for statistical analysis and genetic model building(codominant,dominant,recessive,superdominant,and logadditive models).Then,the correlation between RIG-I and ERCC2 gene polymorphisms and the risk of HBV-HCC in people of different sexes was further analyzed.Statistical methods such as independent samples t-test,analysis of variance,chi-square test,and unconditional logistic regression models were used to detect Hardy-Weinberg equilibrium(HWE),calculate the linkage disequilibrium(LD)and haplotype distribution of each gene fragment,and calculate the odds ratio(OR)and its 95%confidence interval(CI),to comprehensively analyze the relationship between RIG-I and ERCC2 gene polymorphisms and HBV chronic infection and HBV-HCC.Results1.Correlation between RIG-I gene polymorphisms and HBV-HCCBy establishing genetic models,taking the healthy group as the control,and adjusting for age and sex,it can be seen that the rs3824456 G/G genotype and G allele were associated with an increased risk of HBV-HCC(recessive model OR=2.41,95%CI=1.03-5.64;log-additive model OR=1.47,95%CI=1.01-2.14;allele analysis OR=1.45,95%CI=1.01-2.09).However,the rs2777729,rs10813831,and rs3739674 genotypes did not show any significant correlations with HBV-HCC as all P values were greater than0.05 in each genetic model and allele analysis,and there was no significant difference in the results.The stratified analysis also showed that only the G/G genotype at RIG-I rs3824456 was associated with an increased risk of HBV-HCC in the female population(recessive model,OR=6.26,95%CI=1.16-33.88;log-additive model,OR=2.72,95%CI=1.12-6.62).In the male population,no significant association was shown(all P>0.05).The linkage disequilibrium analysis showed that there was a strong linkage disequilibrium between rs3824456,rs2777729,rs10813831,and rs3739674(all D’s were infinitely close to or equal to 1).In haplotype analysis,the top three haplotypes with frequency distributions were CCGG,GAGC,and CAAC,but no haplotype with a significant correlation with HBV-HCC was found(all P>0.05).2.Association of RIG-I gene polymorphisms with HBV chronic infectionBy genetic model analysis,taking the healthy group as the control,after being adjusted by sex and age,it can be seen that the rs10813831 G/A genotype,A/A genotype,and A allele were associated with a decreased risk of chronic HBV infection(codominant model,OR=0.57,95%CI=0.33-0.98;dominant model,OR=0.53,95%CI=0.32-0.89;log-additive model,OR=0.55,95%CI=0.35-0.87;allele analysis,OR=0.53,95%CI=0.33-0.84).However,the genetic models and allele analysis of rs3824456,rs2777729,and rs3739674 did not show any significant correlations with chronic HBV infection(all P>0.05).Linkage disequilibrium analysis showed that there was a strong linkage disequilibrium between RIG-I target SNPs(all D’s were infinitely close to or equal to 1).Haplotype analysis showed that the top three haplotypes with frequency distributions were CCGG,GAGC,and CAAC.Meanwhile,there was an association between the CAAC haplotype and a reduced risk of chronic HBV infection(OR=0.55,95% CI=0.34-0.89).3.Correlation between ERCC2 gene polymorphisms and HBV-HCCThrough the analysis of genetic models,taking the healthy group as the control,after adjusting for age and sex,it can be seen that all genotypes of rs1799785,rs238406,rs238407,and rs238408 had no significant correlation with HBV-HCC(all P values in each genetic model and allele were greater than 0.05).In the stratified analysis,in the female population,recessive models showed that the rs238406 T/T genotype(OR=3.86,95%CI=1.18-12.60),rs238407 T/T genotype(OR=4.18,95%CI=1.27-13.79)and rs238408 C/C genotype(OR=4.18,95%CI=1.27-13.79)were associated with an increased risk of HBV-HCC,while the rs1799785 polymorphism was not significantly associated with the risk of HBV-HCC(P>0.05).However,after stratification,the allele distributions of rs238406,rs238407,and rs238408 in the HBV-HCC group in the female population did not conform to HWE(P<0.05),so this result was for reference only.After stratification,none of the ERCC2 SNP loci showed a correlation with the risk of HBV-HCC in the male population(all P>0.05).The linkage disequilibrium analysis showed that there was a strong linkage disequilibrium between rs1799785,rs238406,rs238407,and rs238408(all D’s were infinitely close to or equal to 1).In haplotype analysis,the top three haplotypes with frequency distributions were TGAG,TTTC,and GTTC,but no correlation with HBV-HCC risk was found(P>0.05).4.Correlation between ERCC2 gene polymorphisms and chronic HBV infectionThrough the genetic model analysis,taking the healthy group as the control,after being adjusted by age and sex,it can be seen that all genotypes of ERCC2 rs1799785,rs238406,rs238407,and rs238408 did not show a significant correlation with HBV chronic infection(all P>0.05).The linkage disequilibrium analysis showed that there was a strong linkage disequilibrium between rs1799785,rs238406,rs238407,and rs238408(all D’s were infinitely close to or equal to 1).In haplotype analysis,the top three haplotypes with frequency distributions were TGAG,TTTC,and GTTC,but there was no significant correlation with chronic HBV infection(P>0.05).Conclusions1.RIG-I rs3824456 gene polymorphisms were associated with HBV-associated hepatocellular carcinoma,and its G allele was associated with an increased risk of HBV-HCC in the Chinese Han population in Jingzhou Prefecture,Hubei Province.There was no significant correlation between rs2777729,rs10813831,and rs3739674 loci polymorphisms and the risk of HBV-HCC.2.RIG-I rs10813831 gene polymorphisms were associated with chronic HBV infection,and its A allele was associated with a decreased risk of chronic HBV infection in the Jingzhou Han population.There was no significant correlation between the gene polymorphisms at the rs3824456,rs2777729,and rs3739674 loci and the risk of chronic HBV infection.3.RIG-I rs3824456,rs2777729,rs10813831,and rs3739674 CAAC haplotype was associated with a decreased risk of chronic HBV infection in the Jingzhou Han population.4.There was no significant correlation between ERCC2 rs1799785,rs238406,rs238407,and rs238408 gene polymorphisms and HBV-HCC and HBV chronic infection in the Jingzhou Han population.5.In the Jingzhou Han female population,ERCC2 rs238406 T/T,rs238407 T/T,and rs238408 C/C genotypes may increase the risk of HBV-HCC,but further studies are still needed.