Screening And Verification Of Single Nucleotide Polymorphisms In ALT And AST Changes Induced By TACE After Resection Of Hepatitis B Virus-related Hepatocellular Carcinoma

Research content:Our current study is divided into five sections:1.Clinicopathological features,prognosis and whole exons analysis of ALT and AST changes induced by TACE after hepatectomy in hepatitis B virus(HBV)-related hepatocellular carcinoma(HCC)patients.2.The second-stage validation cohort of the candidate single nucleotide polymorphism site golgi SNAP receptor complex member 2(GOSR2)-rs 197922 for ALT and AST changes induced by TACE after hepatectomy in HBV-related HCC patients.3.Clinical application value and prospective molecular mechanisms of GOSR2 gene in hepatocellular carcinoma based on genome-wide expression profile dataset.4.Clinicopathological features and prognosis analysis of the expression level of GOSR2 and rs197922 in HBV-related HCC patients.5.Biological function identification of GOSR2 gene in HCC and its relationship between regulation of ALT and AST in normal liver cell lines.Objective:The present study is attempted to identify the whole genome exons polymorphisms of ALT and AST changes induced by TACE after hepatectomy in hepatitis B virus(HBV)-related hepatocellular carcinoma(HCC)patients,and their clinicopathological features and Prognosis.Materials and Methods:A total of 49 patients with HBV-related HCC who underwent hepatectomy in the First Affiliated Hospital of Guangxi Medical University were enrolled.TACE was performed after hepatectomy.Grouped according to changes in ALT and AST after TACE,HCC tumor tissues were used for whole genome exons array scanning and then erformed a whole genome exons association study.Results:We screened 1247 candidate SNPs with a P value<0.05 by performing a whole genome exons association study based on the changes of ALT and AST after TACE.Through ICSNPathway analysis,we identified 17 SNPs(including rs75387493,rs2271683,rs11768465,rs8207,rs28365927,rs1344642,rs1863704,rs10160013,rs4036,rs2235324,rs17183814,rs7973658,rs34550074,rs3740168,rs197922,rs3790525,rs41309181)with altered pathways and found four SNP-Gene-Pathway regulatory mechanisms.Through bioinformatics analysis,we used the GeneMANIA online tool found that the genes of these 17 candidate SNPs have a co-expression interaction relationship with the coding genes of ALT and AST.Through the GTEx database analysis,among these 17 SNPs,we found that golgi SNAP receptor complex member 2(GOSR2)-rs 197922 has a quantitative trait locus relationship in multiple normal tissues such as the normal liver tissue.Conclusions:This study identified 17 potential SNPs with SNP-Gene-Pathway regulatory potential,of which GOSR2-rs 197922 may be involved in the regulation of serum ALT and AST change after TACE.Objective:The present study is an independent second-stage hepatitis B virus(HBV)-related hepatocellular carcinoma(HCC)cohort to verify the results of first-stage GWAS cohort.Materials and Methods:This study collected the tumor tissues from HBV-related HCC patients who underwent hepatectomy in the Department of Hepatobiliary Surgery,the First Affiliated Hospital of Guangxi Medical University from 2014 to 2017,and all of them were underwent TACE after surgery.The genotyping of golgi SNAP receptor complex member 2(GOSR2)-rs197922 was using the Sanger method.Association analysis was performed to reveal the relationship between rs197922 and serum ALT and AST changes before and after TACE.Results:A total of 88 HBV-related HCC patients were enrolled in the second-stage validation cohort.The genotype distribution of rs197922 was as follows:23 AA genotypes,43 AG genotypes and GG.22 cases of genotype.By chi-square test and Ligistic regression analysis,we failed to observe that the rs 197922 genotype distribution was associated with changes of serum ALT and AST after TACE treatment in patients with HBV-related HCC after hepatectomy.Conclusions:Due to changes in the TACE chemotherapy drugs in patients with HBV-related HCC who were included in the second-stage validation cohort,we failed to observe the distribution of rs197922 genotypes were associated with the changes of ALT and AST levels after TACE.Objective:The purpose of current study is to investigated the clinical significance of the golgi SNAP receptor complex member 2(GOSR2)gene based on the genome-wide expression profile dataset of hepatocellular carcinoma(HCC)in GEO and TCGA databases,and investigated the molecular mechanisms in HCC using bioinformatics approaches.Materials and Methods:This study included 212 and 370 hepatocellular carcinoma patients from GSE14520 and TCGA datasets,respectively.The clinical application value of GOSR2 gene was investigated by diagnosing ROC and survival analysis.Then bioinformatics methods such as differentially expressed genes,co-expressed genes and gene set enrichment analysis of GOSR2 were used based on genome-wide expression profile dataset to investigate its potential molecular mechanisms.Results:Through validation of the GSE 14520 and TCGA cohorts,we found that the GOSR2 gene is up-regulation in HCC tumor tissues,and HCC patients with high expression of GOSR2 gene were significantly associated with shorter recurrence-free survival and overall survival time.By comparing the genome-wide expression profile dataset between patients with low-and high-GOSR2 expression groups of HCC,the differentially expressed genes and gene set enrichment analysis between the two groups,and the co-expressed gene of GOSR2 were used for potential molecular mechanisms enrichment.Bioinformatics analysis suggest that GSOR2 may participate in the regulation of HCC biological behavior by regulating cell proliferation,cell cycle,NF-kB and Wnt signaling pathways.Conclusions:GOSR2 may be a potential biomarker for diagnosis and prognosis of HCC.Its biological function may involve the regulation of signaling pathways that such as the cell proliferation and cycle.Objective:To investigate the potential clinical value of golgi SNAP receptor complex member 2(GOSR2)immunohistochemical expression and rs197922 genotype distribution as a biomarker of hepatitis B virus(HBV)-related hepatocellular carcinoma(HCC),and to analyze the Expression Quantitative Trait Loci(eQTL)characteristics of rs 197922 in Guangxi HBV-related HCC.Materials and Methods:Immunohistochemical staining of GOSR2 gene was performed in HBV-related HCC tumor tissues by immunohistochemistry,and rs197922 locus was genotyped.The clinical pathological features and prognosis differences between different GOSR2 gene expression levels and different rs197922 genotypes were compared.Results:This study included 342 patients with HBV-related HCC from Guangxi.Through multivariate survival analysis,we found that the recurrence-free survival and overall survival time between GOSR2 negative and positive immunostaining HBV-related HCC patients were not reach the statistically significant.However,we found that the genotypes of the rs 197922 were significantly associated with the HCC prognosis in subgroup of non-radical resection.Conclusions:In this study,we failed to observe that the immunohistochemistry expression level of GOSR2 gene is significantly associated with the prognosis of HBV-related HCC patients,howe,ver,we observed that the rs 197922 genotype distribution can be used as a potential prognosis biomarker in specific subgroups of HBV-related HCC patients.Objective:The aim of the present study is to investigate the biological function of GOSR2 in hepatocellular carcinoma(HCC)by silencing golgi SNAP receptor complex member 2(GOSR2)gene in HCC cell lines,and to investigate its regulation relationship with ALT and AST in normal liver cell line by treated with the chemotherapeutic drugs of TACE.Materials and Methods:The GOSR2 gene was silenced in HCC cell lines and normal liver cell line by siRNA technique.The biological functions of GOSR2 were verified by Western Blot,RT-PCR,wound healing,proliferation and invasion.The relationship between GOSR2 and ALT and AST in normal liver cell lines was explored by drug induction(5-fluorouracil,pirubicin,cisplatin and lobaplatin).Results:The effect of siRNA silencing GOSR2 in cell lines was verified by WB and RT-PCR.It was confirmed by wound healing,cell proliferation,cell invasion and other experiments that inhibition of GOSR2 gene expression in HCC cell lines can significantly inhibit the cell migration,proliferation and invasion ability.Silencing GOSR2 gene expression in normal liver cell lines,we did not observe significant changes in the secretion of ALT and AST from normal liver cell lines after chemotherapy drugs induction.Conclusions:This study confirmed that the GOSR2 gene may play an oncogene role in HCC,but the present study did not observe a direct regulation relationship between GOSR2 and ALT and AST after chemotherapy drugs induction in normal liver cell line.