Primary Hepatocellular Carcinoma And Th1/Th2Cytokines Gene Polymorphism

Background Hepatocellular carcinoma (HCC) is a malignant tumor. Chronic hepatitis B virus (HBV) infection is an important risk factor. But the individuals had the different immune response and clinical outcome to HBV infection. Some of the cytokines single nucleotide polymorphisms (SNP) influence the immune function, HBV infection outcomes and HCC occurrence.This research was divided into two parts to explore the association of cytokine SNP with HCC. The first part discussed the association of Thl cytokines IL-2, IFN-y gene polymorphism with HBV infection and HBV related HCC. Meanwhile, to explore the interaction of HCC risk factors with cytokine gene. The second part combined the cytokines IL-2, IFN-γ, IL-1B of Th1and IL-6, IL-10of Th2, to explore the association of Thl/Th2cytokine SNP with HCC. The purpose of this study was to provide the reference for HCC risk assessment, prevention and treatment. PART1THE ASSOCIATION OF TH1CYTOKINE IL-2AND IFN-GAMMA GENE POLYMORPHISM WITH HBV INFECTION AND HBV RELATED HCCObjective To explore the cytokines IL-2gene and IFN-gamma gene single nucleotide polymorphisms in Guangxi people, and the impact of HBV infection and HBV-HCC occurrence. To analyze the interaction of gene with environment factors such as smoking, alcohol using and HCC related family history, in HBV infection and HBV-HCC occurrence.Method A case-control study based on hospital was carried out and all the objects were frequency matched by375HCC patients-377HBV carriers-406normal control. All the subjects were recruited from the First Affiliated Hospital of Guangxi Medical University, Tumor Hospital of Guangxi and the People’s Hospital of Guangxi during2007June-2010July. All the subjects were surveyed face to face by the specifically trained investigators, including basic datum and environment exposure factors. Clinic data were add by medical records. TaqMan MGB Real-Time fluorescence quantitative PCR technology based on ABI platform was applied to detect the SNPs of IL-2-33OA/C(rs2069762), IFN-γ-1615GG/A(rs2069705) and+5171T/C (rs2069727). All data were input and analyzed statistically in SPSS16.0for Windows. The interaction of gene-environment in the three groups were analyzed by Logistic regression model and assessed by the synergy index S. The Hardy-Weinberg equilibrium and linkage disequilibrium of IFN-gamma haplotype were analyzed in HaploView4.2. Result (1) A total of1158subjects with complete information were recruited,375for HBV-HCC patients,377for HBV carriers, and406for normal controls. There were no statistically differences in the distributions of age, gender, nation and marriage, but significant statistically difference in environment factors smoking, alcohol using and HCC related family history (P<0.05). The risk of HBV-HCC in normal control who had smoke, alcohol use and family history were2.12(1.32-3.40)、3.41(2.14-5.42)、22.63(8.97-57.10) respectively, and in HBV carriers were1.52(0.97-2.38)、2.25(1.45-3.50)、41.34(12.80-133.45) respectively.(2) The damage of liver was serious in HBV-HCC patients. The indexes of liver function in HBV-HCC patients were significant different with HBV carriers and normal controls except TA. The damage of liver was lighter in HBV carriers, there were statistically differences in ALB, ALT and ALP compared with normal controls.(3) There were no significant statistically difference in the polymorphisms of IL-2-330A/C, IFN-γ-1615G/A and+5171T/C among the three groups (P>0.05).(4)-1615C/T and+5171A/G sites on IFN-gamma had linkage disequilibrium(D’=0.976, P=2.22-16). The frequency of haplotype GC in population was less the0.03. The rest haplotypes between HBV-HCC groups and the total controls (HBV carriers and normal control) had no significant statistically difference. (5) The interaction of smoking with IL-2and IFN-gamma gene were negative in HBV infection and HB V-HCC occurrence (S<1).The interaction of alcohol using with IL-2-330A/C was positive. Alcohol using combined IL-2-330C increased the risk of HBV infection and HB V-HCC occurrence. The interaction of alcohol using with IFN-y-1615G/A and+5171T/C was negative in HB V-HCC occurrence, and positive in HBV infection. Alcohol using combined IFN-y-1615A and IFN-y+5171C increased HBV infection risk.No matter carried HBV or not, someone who had HCC related family history combined IL-2-330loci, IFN-y-1615or IFN-γ+5171loci could inhence HB V-HCC occurrence.Conclusion (1) There was no significant statistically difference in the polymorphisms of IL-2-330A/C, IFN-γ-1615G/A and IFN-γ+5171T/C among HB V-HCC patients, HBV carriers and normal controls.(2) Smoking, alcohol using and HCC related family history are the important risks of HBC-HCC.(3) There were interaction of these3loci polymorphisms with environment exposure factors. The mutant genes might not influence the occurrence of HCC and HBV infection directly in the population of Guangxi, but enhance the risk interacted with the environment risk factors. PART2THE ASSOCIATION OF TH1/TH2MAIN CYTOKINES GENE POLYMORPHISM AND HEPATOCELLULAR CARCINOMAObjective According to the SNP distribution of Thl and Th2cytokines SNP in Guangxi people, to explore the association of Th1/Th2cytokines gene-gene combined effect with HCC occurrence.Methods A case-control study based on hospital was carried out.720HCC patients from Guangxi Medical University first affiliated hospital and Tumor Hospital of Guangxi during2007June-2010July and848controls from physical examination center of Guangxi Medical University first affiliated hospital and Guangxi People’s hospital during the same period were investigated with environmental exposure questionnaire. Clinic data were add by medical recors. TaqMan fluorescence quantitative PCR technology based on ABI platform was applied to detect the SNPs of Th1cytokines IL-2-33A/C(rs2069762), IFN-γ-1615G/A (rs2069705) and+5171T/C(rs2069727), IL1B-31A>G (rs1143627) and-511G>A (rs16944), and Th2cytokines IL-6-572C>G (rs1800796), IL-10-592A>G (rs1800871) and-819T>G (rs1800872). All data were input and analyzed statistically in SPSS16.0for Windows. Measurement data were compared with the use of the Student’s test. While categorical data were compared with the use of the chi-square test. Non-conditional logistic regress model was used for relative risk scale, indicated as Odd ratio and95%confidence interval indicate. HaploView4.2was used for Hardy-Weinberg equilibrium test. Results (1) A total of1568subjects with complete information were recruited,720for HCC and848for controls. There were no statistically differences in the distributions of age, gender and nation. Gender is the risk factor of HCC, the risk in male is1.54times than female. The environment factors smoking, alcohol using, HCC related family history and HBV infection are the risk factors, OR(95%CI) were2.10(1.49-2.96),2.52(1.79-3.54),25.82(11.63-57.34),5.96(4.60-7.74) respectively.(2)There was no significant statistically difference in the polymorphisms of IL-2-330A/C(rs2069762), IFN-γ-1615G/A (rs2069705) and+5171T/C(rs2069727),IL1B-31A>G(rs1143627)and-511G>A(rs16944),IL-6-572C>G (rs1800796), IL-10-592A>G (rs1800871) and-819T>G (rs1800872) between HCC and cancer free controls (P>0.05).(3)Th1and Th2risk genotype carried number were significantly different in case and control. The risk of HCC increased with the numbers of risk genotypes of Th1and Th2. Individuals with the maximum number(4-5) of Thl genes putative risk genotypes exhibited an OR of6.14(95%CI,3.17-11.92) compared to minimum number(0-1), and middle number(2-3) exhibited an OR of4.33(95%CI,2.67-7.02). While, The risk of carrying2-3Th2risk genotypes was1.51times than the minimum carrier(95%CI:0.96～2.37).(4)There were significantly differences in Th1and Th2risk genotypes between case and control in minority Zhuang group, alcohol using people and no family history people, and HCC risk increased with the number of Thl and Th2risk genotypes(P trend<0.05). There were significantly different in Thl risk genotypes between case and control in male, nation Han, no smoking or alcohol using habit people and HBsAg+patients, HCC risk increased with the carried number of Thl risk genotypes(Ptrend<0.05). No matter smoking or infected HBV, HCC risk increased with the number of Th1risk genotypes(P trend<0.05).(5)Compared with carrying minimum risk genotype in both Thl and Th2, the more risk genotypes were carried in individuals, the higher risk of HCC was showed (P trends<0.05). The risk of HCC in the individuals who carried4-5Thl risk genotype meanwhile2-3Th2risk genotype, was23.42times than the0-1Th1-Th2carrier.Conclusion (1)there were no significantly difference in SNPs of Thl cytokines IL-2-33OA/C(rs2069762), IFN-γ-1615G/A(rs2069705) and+5171T/C (rs2069727), IL1B-31A>G (rs1143627) and-511G>A (rs16944), and Th2cytokines IL-6-572C>G (rs1800796), IL-10-592A>G (r1l800871) and-819T>G (rs1800872) between Guangxi HCC patients and healthy people.(2)Thl and Th2risk genotype carried number were significantly different in case and control. The risk of HCC might be increased with the numbers of risk genotypes of Th1and Th2. Thl-Th2combined effect might enhance the risk of HCC.