Study On The Relationship Between Apoptotic Protease Activating Facter-1 Gene Single Nucleotide Polymorphism And Susceptibility To Hepatocellular Carcinoma In Liver Cancer Famliy Pedigree In Guangxi

Objective:1.To investigate the significance of APAF1 protein expression in the hepatocellular carcinoma.2. To study the relationship between the apoptotic protease activating facter-1(APAF1) gene’s single gene nucleotide polymorphisms (SNPs) of rs2118852 and rs2278361 and genetic susceptibility to hepatocellular carcinoma (HCC) in liver cancer families groups in Guangxi.Methods:1.50 cases of primary hepatocellular carcinoma specimen,50 cases of carcinoma adjacent specimen and 20 cases of normal liver specimen (control group) were selected in this test. The immunohistochemistry methods (streptavidin-perosidase S-P) were established for detecting the APAF1 protein expression in hepatocellular carcinoma tissues, carcinoma adjacent tissues and normal hepatocellular tissues, and chi-square test was carried out to analyze the difference in APAF1 protein positive rate among different groups. 2. In the case control study,20 liver cancer family groups (78 members) as the case and 10 normal family groups (40 members) as the control were involved. Time of Flight Mass Spectrometer (TOF) was used to detect the genotype frequencies of APAF1 SNP of rs2118852 and rs2278361 polymorphisms, and non-conditional logistic regression was carried out to analyze the correlation between the gene polymorphism and HCC risk.Results:1. The positive expression of APAF1 protein in HCC tissues, adjacent tissues and normal liver tissues were 21 (50 cases),39 cases (50 cases) and 19 cases (20 cases), and the positive rates were 42%,78% and 95%, respectively, P<0.05, and the difference was statistically significant. The positive rate of APAFlprotein in HCC was related to the age, sex, tumor size, and hepatitis B and AFP (P>0.05)2.(1) The frequencies of the APAF1 gene rs2118852 genotypes GG, GT and TT were 62.82%,33.3% and 3.85% among the liver family group. The frequencies of the APAF1 gene rs2118852 genotypes GG, GT and TT were 82.05%,17.95% and 0.00% among the normal controls. Allele distributions did not differ significantly between two groups (P>0.05), and genotype distributions conformed to the Hardy-Weinberg equilibrium.(2) In the normal controls,the odds ration of HCC for members with GT genotype was 1.58 (95%CI=0.28～8.89, P= 0.401>0.05),and the difference was not statistically significant.The odds ration of HCC in the normal control group with the TT genotype couldn’t be calculated. In the HCC high incidence families groups, the odds ration of HCC for members with GT and TT genotypes were 0.50(95%CI=0.13～2.00, P=0.322>0.05) and 12.36(95%CI=0.80-190.80, P=0.072>0.05),and the difference was not statistically significant.3.(1) The frequencies of the APAF1 gene rs2278361 genotypes TT, TC and CC were 71.79%,28.21% and 0.00% among the liver family group. The frequencies of the APAF1 gene rs2278361 genotypes TT, TC and CC were 69.23%,25.64% and 5.13% among the normal controls. Allele distributions did not differ significantly between two groups (P>0.05), and genotype distributions conformed to Hardy-Weinberg equilibrium.(2)In the normal controls,the odds ration of HCC for members with TC genotype was 0.68(95%CI=0.14～3.34, P=0.638>0.05),and the difference was not statistically significant.The odds ration of HCC in the normal control group with the CC genotype couldn’t be calculated. In the HCC high incidence families groups, the odds ration of HCC for members with TC genotype was 0.865(95%CI=0.13～2.00, P=0.838>0.05),and the difference was not statistically significant.The odds ration of HCC in in the HCC high incidence families groups with the CC genotype couldn’t be calculated.Conclusion:1-. There is an abnormal expression of APAF1 gene in patients with hepatocellular carcinoma, and its protein expression was down regulated in hepatocellular carcinoma tissues.2. Genotype distributions of APAF1 gene rs2118852 and rs2278361 in both group people conformed to the Hardy-Weinberg equilibrium.3. The APAF1 gene rs2118852 and rs2278361 polymorphisms may not be correlated with liver cancer in family clustering genetic susceptibility in Guangxi.