The Relationship Between ERCC1 And AFP Single Nucleotide Polymorphisms And Hepatocellular Carcinoma

Objective1.To explore excision repair cross complementing 1(ERCC1)rs735482,rs1046282,rs3212948 three sites and alpha-fetoprotein(AFP)rs4024,rs737241 two sites of single nucleotides explore the relationship between the susceptibility of single nucleotide polymorphisms(SNPs)and hepatocellular carcinoma(HCC)and the clinicopathological features of HCC.2.To investigate the relationship between ERCC1 mRNA expression,AFP mRNA expression and gene polymorphism and clinicopathological features of HCC.3.To explore the relationship between ERCC1 protein level,serum AFP level and gene polymorphism and clinicopathological features of HCC.METHODSA total of 118 new HCC patients who were not treated with radio-chemotherapy and surgery were enrolled in the case group.The control group received 88 patients with no liver lesions and tumors medical examination staff(age,gender was mathed with HCC group).Snapshottechnique was used for genotyping the SNP sites of the ERCC1 and AFP genotypes,the relationship between ERCC1 and AFP gene polymorphisms and the risk of HCC risk for odds ratio,(OR)and 95% confidence interval(CI)was performed.Real-time quantitative PCR(Qrt-PCR)was determined to confirm for gene expresion.The level of serum AFP was detected by chem-iluminescence immunoassay.The expression of ERCC1 protein was detected by immunohistochemistry.The clinic-pathological features of patients with HCC were collected and the relationship between these clinic-pathological features and ERCC1 and AFP gene polymorphisms was analyzed.The Hardy Weinberg equilibrium law test(HWE)is used to determine whether the genotype distribution of the subject is representative.The test used to analyze the risk of genotype and liver cancer,calculate the OR value and 95% CI,and unconditional logistic regression method statistical analysis to correct the influence of confounding factors such as gender,age,drinking history and smoking history on OR value and 95% CI.The relationship between clinicopathological features and ERCC1 and AFP gene polymorphisms was analyzed by rank sum test.The haplotype model was used to analyze the gene interaction using SHEsis.Statistical analysis was performed using SPSS 17.0,all the P-values were 2-sided,statistical significance was defined as P<0.05.RESULTS1.There was no significant difference in the frequency of allele and genotype distribution between ERCC1 gene rs735482,rs1046282 and rs3212948 in the case group and control group(both P>0.05),suggesting that the three sites of ERCC1 and liver cancer have not been found sensually relevant.The ERCC1 locus polymorphism and clinicopathological analysis showed that the C allele of the 1046282 mutation was associated with increasedexpression of AFP(P = 0.011),and the mutation of the T allele at the rs3212948 locus may affect Pathological grade of liver cancer(Edmondson-Steiner,EDSON)(P = 0.022),carrying the C allele of rs1046282 locus may be associated with decreased DNA load of hepatocellular carcinoma(P = 0.035),carrying the rs735482 locus A allele may be associated with The diameter of the hepatocarcinoma tumor was increased(P = 0.037).2.In 50 HCC tissues,the expression level of ERCC1 mRNA in cancer tissues was 1.158(0.826,2.958),and the expression level of ERCC1 mRNA in paracancerous tissues was 1.230(0.318,3.159).The ERCC1 mRNA level and the tumor size of HCC and tumor thrombus The clinicopathological features of liver cancer such as invasion and intact capsule were not correlated(P>0.05).The rs735482,rs1046282,rs3212948 polymorphisms were not associated with the expression of ERCC1 mRNA in liver cancer(all P>0.05).3.There were 22 cases(22/65,33.85%)of ERCC1 protein positive in HCC tissues of 65 cases.The results showed that ERCC1 rs735482,rs1046282,rs3212948 polymorphisms were not associated with ERCC1 protein expression in HCC tissues(all P>0.05).The clinicopathological analysis of ERCC1 protein and tumor size,tumor invasion,and capsule integrity suggest that ERCC1 protein expression is not associated with clinicopathological factors such as tumor size,tumor invasion,and intact capsule(all P>0.05).4.The genotype frequencies of AFP gene rs737241 in the case group were:GG was 59(50.00%),GA was 51(43.20%),AA was 8(6.78%);control group genotype frequency distribution: GG was 31(35.20%),GA was 42(47.70%),AA was 15(17.70%),and the frequency distribution of GG,GA and AA genotypes was statistically significant(P=0.023).Rs737241 locus polymorphism the risk of HCC in individuals carrying the A allele was lowerthan that of individuals carrying the G allele,and the A allele was 0.573 times higher than the G allele(P = 0.008,OR = 0.573,95%CI 0.3795-0.865);individuals with at least one mutant allele genotype(GA+AA)were 0.543 times more likely to have HCC than individuals carrying the GG genotype(P = 0.037,OR = 0.543,95% CI 0.306-0.965);individuals carrying the homozygous mutant AA genotype were 0.297 times more likely to develop HCC than individuals carrying the wild-type GG gene(P = 0.014,OR = 0.297,95% CI 0.112-0.785).Patients with AA haplotypes carrying the AFP gene rs737241-rs4024 were associated with a significant reduction in HCC risk(P = 0.044,OR = 0.393,95%CI 0.155-0.997),whereas GG haplotypes were associated with a significant increase in risk of liver cancer(P=0.017,OR=2.130,95%CI 1.135-3.999).The rs4024 polymorphism was not associated with the susceptibility of liver cancer(all P>0.05).AFP locus polymorphism and clinicopathological analysis showed that the A allele carrying rs737241 mutation may be associated with elevated serum AFP levels(P = 0.007),and the rs4024 locus carrying the mutant A allele may It was associated with a reduced risk of liver cancer vascular invasion(P=0.013).5.The expression level of AFP mRNA in 50 cases of liver cancer was1.868(0.826,17.195),and the expression level of AFP mRNA in adjacent tissues was 1.079(0.287,3.159).The expression of AFP mRNA was correlated with the Barcelona Clinic Liver Cancer(BCLC)(P=0.006),and was not associated with pathological parameters such as tumor diameter,EDSON grade,and tumor thrombus(all P>0.05).The polymorphisms of rs4024 and rs737241 were not found to be associated with the expression of AFP mRNA in hepatocellular carcinoma(all P>0.05).6.The serum AFP levels of rs4024 locus and rs737241 locus GA+AAgenotype and GG genotype were compared,and there was no significant difference in serum AFP levels between the two groups(P>0.05).The serum AFP level of HCC was correlated with the age,BCLC stage and EDSON grade of HCC patients(P<0.05).Further multivariate analysis showed that the smaller the age of liver cancer patients,in the 118 case the higher the serum AFP level(P = 0.001,OR=0.172,95% CI 0.059-0.501);the higher the BCLC stage of liver cancer patients,the higher the serum AFP(P = 0.000,OR = 4.420,95% CI2.003-9.752);the higher the EDSON grade of liver cancer patients,The higher the serum AFP level(P = 0.034,OR = 2.335,95% CI 1.068-5.102).CONCLUSION1.The ERCC1(rs1046282,rs3212948,rs735482)locus polymorphism was associated with AFP levels,tumor size,and HBV DNA load pathological parameters of HCC.The ERCC1 gene polymorphism has not been found to be associated with susceptibility to hepatocellular carcinoma.2.The ERCC1 gene polymorphism has not been found to be associated with ERCC1 mRNA and protein expression.3.There was a certain correlation between the AFP gene rs737241 polymorphism and the risk of HCC.The AFP(rs4024,rs737241)gene polymorphism was associated with HCC tumor thrombus and serum AFP levels.4.AFP mRNA expression may be associated with BCLC staging of HCC.High expression of AFP mRNA may be associated with clinical progression of HCC.