Associations Of FOXO Gene Snps With Hepato-Cellular Carcinoma Clinico-pathological Features Ang Prognosis And With The Expression Of FOXO Protein

ObjectivesHepatocellular carcinoma(HCC)is a high malignancy with insidious onset,and its prognosis is very poor,but we know little about the main factor affecting features and outcome at present.Growing bodies of evidences indicate that forkhead transcription factor box O(FOXO)plays an important role in tumor progression and prognosis,but FOXO genes have any influence to the clinico-pathological characteristics and prognosis in HCC not yet to know.So in this study we are trying to explore the relationships between FOXO gene SNPs and the clinical pathological characteristics and prognosis of HCC,and also detect the association between positive locus and corresponding FOXO protein expression in plasma,aimed to provide referenced evidences for the prevention and prognosis of HCC evaluation.MethodsThe current study patients were those who were diagnosed as HCC the first time and were hospitalized patients in Guangxi medical university first affiliated hospital and affiliated tumor hospital.The basic information of all the participants was collected by field epidemiological investigation;the clinico-pathological characteristics and the treatment process were collected from patient medical records.Participants were followed-up to observe tumor recurrence,metastasis and death or not after radical local hepatectomy by telephone follow-up and reviewing medical records.We screened the SNPs of FOXO genes in the China population on America National Institute of Environmental Health Sciences database and selected the high frequency of mutations for this study which the minimum allele frequency above 0.05.Then we got three SNP locus,there were rs17592236(C>T)of FOXO1 gene,rs4946936(C>T)of FOX03 gene and rs4503258(C>T)of FOX04 gene.And high flux TaqMan MGB real-time fluorescent quantitative PCR technology was applied on genotyping these three SNPs.FOXO protein concentrations in plasma was measured by ELISA method.The datas were double inputed and validated with the Epidata 3.1 software and SPSS 17.0 software was using for statistical analysis.Chi-square test or exact test was used to explore the association between SNPs and clinico-pathological characteristics of HCC,the odds ratio(OR)and 95%confidence interval(CI)were calculated by unconditional logistic regression model.When analysing the survival,Kaplan-Meier method was used to univariate analysis,and Cox proportional hazards model was used for multivariate analysis.All statistical tests were two-sided,a=0.05.Results(1)The associations between SNPs of FOXO gene and clinic-pathological features in HCC.Chi-square test results showed that the polymorphism of rs17592236 associated with tumor thrombus(?2=7.59,P=0.022)and vascular invasion(?2=7.08,P=0.029),the genotype TT increased the risk of appearing vessel invasion(OR=3.926,95%CI:1.333?11.568)and genotypes(CT+TT)had a greater risk of cancer embolus(OR=1.271,95%CI:1.065?1.518)after controlling for confounders.The polymorphism of rs4946936 was related to vascular invasion(?2=4.82,P=0.028)too,and genotypes(CT+TT)decreased the risk of appearing vessel invasion(OR=0.788,95%CI:0.642?0.996).There was no statistical difference between other clinic-pathological features and the polymorphisms of rs4946936 and rs17592236,and there was not statistically significant between the polymorphism of rs4503258 and all clinic-pathological features(all p>0.05).(2)The relationships between SNPs of FOXO gene and prognosis in HCCThe HCC 5-years accumulate survival rate was 55.8%and 5-years progression-free survival rate was 35.3%after radical local hepatectomy.K-M univariate analysis results showed that tumor progression was correlated with polymorphisms of rs4946936 and rs17592236(?2 =7.7,P=0.005 and ?2 =5.7,P=0.017),genotype TT of both these two SNPs increased the risks for tumour progression(HR=2.325,95%CI:1.295-4.174 and HR=1.659,95%CI:1.042?2.643)and their PFS(progression-free survival)was short.Analysis results also showed that the risk of death was closely related to polymorphism of rs4946936(?2 =4.6,P=0.031),the homozygous of the rs4946936 variant allele TT increased the risk of death(HR=2.006,95%CI:1.017?3.956)and enjoyed shorter OS(overall survival time).The risk of death was not correlation with rs17592236 polymorphism and the polymorphism of rs4503258 was irrelevant to HCC progression or death(all p>0.05).Analysis results also showed that the risks of progression and death were higher in BCLC-(B+C)stages(Barcelona stage)(HR=1.51,95%CI:1.023?2.231 and HR=1.861,95%CI:1.152?3.004)and their OS and PFS were short.AFP(alpha fetoprotein)concentrations greater than 1000 ng/mL increased the risk of progression and death(HR=1.593,95%CI:1.067?2.378 and HR=1.628,95%CI:1.010?2.623)and had shorter OS and PFS.Both comprehensive treatment and tumor cellular differentiation had no association with HCC progression or death(allp>0.05).(3)The correlations between polymorphism of FOXO3 gene and FOX03 protein concentrations in plasma.The average concentrations of FOX03 protein were(17.53±9.69)ng/mL in plasma,and there was no statistical significance between FOXO3 protein concentrations and the polymorphism of rs4946936.Conclusion(1)The genotype TT of rs17592236 increased the risk of vascular invasion and the mutant T allele increased the risk of cancer embolus while the mutant T allele of rs4946936 reduced the risk of vascular invasion.(2)The genotype TT of rs4946936 increased of risk for both tumour progression and death;its PFS and OS were short.The genotype TT of rs17592236 decreased the risk of tumour progression and had shorter PFS.Patients who were BCLC-(B+C)stage or AFP?1000ng/mL had higher risk for tumour progression and death.(3)There was no relationship between FOXO3 protein concentrations and the polymorphism of rs4946936.On the whole,the polymorphisms of FOXO genes had close relationships with the clinic-pathologic features and prognosis of HCC.