Associaton Of Several Non-immune Related Gene Polymorphisms With The Susceptibility Of HBV-related Heaptic Cirrhosis

Background and Objective: There are different clinical outcomes in patients with hepatitis B virus infection, and the most serious condition is hepatic cirrhosis. The development of HBV-related hepatic cirrhosis is associated with not only environmental and viral factors but also hereditary susceptibility. Single nucleotide polymorphisms (SNPs), account for more than 90% of human genetic variations, are the best genetic markers to indicate genetic susceptibility of diseases. There are some studies reported on the relationships between cytokine gene polymorphisms and the HBV-related hepatic cirrhosis, but few studies on the non-immune-related gene polymorphisms associated with the development of HBV-related cirrhosis, and lack of systematic researches about them. In the present study, we detected 7 genetic polymorphism sites of non-immune-related genes possiblely associated with the development of liver cirrhosis, and analyzed the relationship between them and the susceptibility of HBV-related hepatic cirrhosis.Methods: (1) Patients: 168 cirrhosis patients with HBV infection (case group) and 155 asymptomatic HBV carriers (control group) were recruited. (2) Gene polymorphism analysis: peripheral blood genomic DNA was extracted by phenol-chloroform method. Polymerase chain reaction-restriction fragment length polymorphism was used to determine the genotypes of each polymorphism sites. (3) Genotype and allele frequencies in each site were calculated and their differences between two groups were compared by chi-square test, and Hardy-Weinberg equilibriums of genotypes was verified by chi-square test. (4) The odds ratios (ORs) and their 95% confidence intervals of all genotypes and alleles were calculated by univariate Logistic regression analysis in total sample and gender-stratified sub-samples, respectively. Multivariate Logistic regression analysis was used to screen independent risk and protective genotypes and alleles.Results: (1) The differences of all genotype and allele frequencies at AGT-20A/C site were not significant between case group and control group (P>0.10). However, multivariate analysis showed a probable association between this site and susceptibility of HBV-related cirrhosis in female patients, the ORs of genotype AA and allele C nearly significant (P=0.064, 0.080). (2) The differences of all genotype and allele frequencies at AGT -6A/G site were not significant between case group and control group (P>0.10), but multivariate analysis showed that AA was a risk genotype (OR=1.927, P=0.047) and G was protective allele (OR=0.508, P=0.047) in male patients. (3) The differences of all genotype and allele frequencies at ESR1 +29T/C site were not significant between case group and control group (P>0.10). However, univariate analysis showed that it is closely associated with susceptibility of female cirrhosis. The TC was susceptible genotype (OR=3.336, P=0.007), CC was a protective genotype (OR=0.327, P=0.015), and T was susceptible allele (OR=3.061, P=0.015) in female patients. Multivariate analysis confirmed that TC was susceptible genotype (OR=3.454,P=0.027) and T was susceptible allele (OR=3.554,P=0.024) in female. (4) The differences of all genotype and allele frequencies at MMP9 -1562C/T site were not significant between case group and control group (P>0.10), and univariate and multivariate analyses did not show association between it and susceptibility of HBV-related cirrhosis. (5) The differences of all genotype and allele frequencies at MEH Tyr113His site were not significant between case group and control group (P>0.10), and univariate and multivariate analyses did not show association between it and susceptibility of HBV-related cirrhosis. (6) The differences of all genotype and allele frequencies at MEH -613C/T site were not significant between case group and control group (P>0.10), but there seemed some correlation between this site and susceptibility of female HBV-related liver cirrhosis. In female patients, the ORs of TT genotype and C allele were nearly significant (P=0.089, 0.089). (7) The differences of GSTM1 genotype frequencies were not significant between case group and control group (P>0.10), and univariate and multivariate analyses did not show that this site is associated with susceptibility of HBV-related liver cirrhosis. (8) Multivariate Logistic regression analysis showed that in the 7 genetic variations detected, AGT-6AA was an independent risk genotype and AGT-6G was an independent protective allele for HBV-related cirrhosis development in male patients, and both of ESR1+29TC genotype and T allele were risk factors for HBV-related cirrhosis development in female patients.Conclusions: (1) AGT gene polymorphisms are associated with susceptibility of HBV-related liver cirrhosis, in which -6A/G site correlated with susceptibility of male cirrhosis, and -20A/C correlated with susceptibility of female cirrhosis. (2) There is significant association between gene polymorphisms at position +29T/C of ER1 and susceptibility of female HBV-related liver cirrhosis, but without significant association with male. (3) There is no significant association between gene polymorphisms at position -1562 of MMP-9 and susceptibility of HBV-related liver cirrhosis. (4) There is no significant association between gene polymorphisms at position -613C/T or -Tyr113His of MEH and susceptibility of HBV-related liver cirrhosis. (5) There is no significant association between null or normal genotype of GSTM1 and susceptibility of HBV-related liver. (6) AGT-6A/G site is an independent factor for male cirrhosis development and ESR1+29T/C site is an independent factor for female cirrhosis development.