Study Of Novel Biomarker EPRS In Hepatocellular Carcinoma

Although there are many therapeutic methods and drugs for hepatocellular carcinoma(HCC),the overall therapeutic effect of HCC is still poor due to low early diagnosis rate,poor liver function status of patients and poor drug tolerance.Future treatment of HCC should target multiple pathways in HCC and adopt personalized therapy with combination of multiple targeted drugs to improve overall survival.Therefore,the search for more precise molecular diagnostic and prognostic markers is the trend of accurate therapy for HCC.In this paper,integrated bioinformatics methods are used to screen out novel markers for diagnosis and prognosis of HCC and verify them,thus exploring a new idea for drug development of targeted therapy for HCC.The study consists of three parts:Part 1.Screening of hepatocellular carcinoma biomarkers by integrating genomics and proteomics.Objective:To screen the diagnostic and prognostic markers of hepatocellular carcinoma using publicly available genomic and proteomic data.Methods:1.Bioinformatics methods were used to screen novel markers for HCC diagnosis and prognosis based on publicly available genomic and proteomic databases.2.Fresh cancer tissues and adjacent tissues of HCC patients were collected for preliminary verification using RT-qPCR and Western blot respectively.Results:1.EPRS was selected as the candidate research target based on bioinformatics method.2.The mRNA(p=0.0401)and protein(p=0.0484)expressions of EPRS in HCC tissues were higher than those in adjacent normal tissues.Conclusion:EPRS can be used as a candidate marker for HCC.Part 2.Expression significance of EPRS in hepatocellular carcinoma Objective:To investigate the relationship between EPRS expression and clinicopathological features and survival.Methods:1.After downloading and integrating the mRNA expression profile data and clinicopathological data of hepatocellular carcinoma from TCGA database,the relationship between EPRS mRNA and clinicopathological features and prognosis was analyzed.2.The expression of EPRS protein in HCC tissue microarray was detected by immunohistochemistry,and the relationship between EPRS and clinicopathological features and prognosis was analyzed.Results:1.EPRS mRNA was significantly correlated with TNM stage(p=0.0001),histological grade(p=0.0002),tumor size(p=0.001)and vascular invasion(p=0.0123).2.Patients with high EPRS mRNA expression had shorter overall survival than those with EPRS mRNA expression(p=0.0004).3.EPRS mRNA(p=0.025)was an independent adverse prognostic factor affecting hepatocellular carcinoma survival.4.EPRS protein was significantly correlated with tumor size(p=0.029),portal vein embolism(p=0.024)and TNM stage(p=0.0315).5.Patients with high expression of EPRS protein had shorter overall survival(p=0.0003)and disease-free survival(p=0.0076)than those with low expression of EPRS protein.6.Univariate and multivariate COX survival analysis showed that EPRS protein was an independent adverse prognostic factor affecting the survival of HCC patients(p=0.047)and relapse-free survival(p=0.009).Conclusion:EPRS can be a potential prognostic marker for HCC.Part 3.Study on the relationship between EPRS and HepG2 cell proliferation,apoptosis and invasion abilityObjective:To explore the relationship between EPRS and hepatocellular carcinoma proliferation,apoptosis and invasion ability.Methods:1.Construct stable transfected cells with lentiviral plasmid that interferes or enhances EPRS gene expression in HepG2 cell lines.2.The proliferation,invasion and apoptosis of stable transfected cells in each group were detected in vitro,and xenograft tumor models were constructed to further verify the effect of EPRS on the proliferation of HepG2 cells in vivo.Results:Proliferation capacity and invasion capacity of HepG2 cell lines in EPRS overexpression group were higher than those in normal control group(p=0.0253),proliferation capacity and invasion capacity of HepG2 cell lines in EPRS knockdown expression group were lower than those in normal control group(p=0.0421),but there was no statistical difference in apoptosis rate among the three groups.The results of subcutaneous tumor formation in nude mice showed that cell proliferation rate(p=0.0351)and weight(p=0.035)in EPRS overexpression group were higher than those in EPRS knockdown expression group.Conclusion:EPRS can promote the proliferation and invasion of hepatocellular carcinoma.