Preliminary Study On Regulation Of Des-γ-carboxy Prothrombin Expression By Long Non-coding RNA ATB In HBV-associated Hepatocellular Carcinoma

Objective: Hepatocellular Carcinoma(HCC)is one of the most common malignant tumors of digestive system,which ranks 6th and 4th in incidence and mortality in the world respectively.Tumor markers play an important role in the early diagnosis,screening and prognosis of HCC.As an important tumor marker,Des-γ-carboxy prothrombin(DCP)has been used in the clinical diagnosis,screening and prognosis judgement of HCC.DCP plays an important role in the carcinogenesis of HCC,but the mechanism of DCP expression regulation has not been reported in the literature.Hepatitis B virus(HBV)infection is a high risk factor for the development of HCC,accounting for 54% of all HCC patients worldwide and more than 80% of all Chinese Hepatitis B virus.However,the exact mechanism of how HBV infection leads to HCC remains unclear.Our previous experimental study found that in patients with HBVassociated HCC,serum DCP levels were significantly higher in the group with high viral load(HBV-DNA ≥2000 IU/m L)than in the group with low viral load(HBVDNA < 2000 IU/m L);The level of serum DCP in the anti-virus treatment group was significantly lower than that in the non-treatment group,and there was a positive correlation between the level of serum DCP and HBV-DNA load.It is suggested that the tumor marker DCP is closely related to HBV in HBV-related HCC,but there is no related research report so far.Long non-coding RNAs(Lnc RNAs)are non-coding RNAs with a length of more than 200 nucleotides.lnc RNA plays an important role in the development of many kinds of tumors,including liver cancer,colon cancer,stomach cancer and lung cancer.Recent studies have shown that lnc RNAs are involved in the initiation and progression of HCC through a variety of regulatory mechanisms.The aim of this study was to investigate the effect of HBV-related lnc RNAs on the regulation of DCP expression in HBV-associated HCC.This study provides a new molecular target and theoretical basis for the regulation of DCP expression in HCC and its clinical application.Methods:1.To screen out lnc RNAs related to HBV-DNA viral load through TCGA database,related literature and differential analysis of lnc RNAs expression level in serum of HBV-related HCC patients;2.Using real-time quantitative PCR to screen lnc RNAs related to DCP concentration in patients with HBV-related HCC.3.In PLC/PRF/5 hepatocellular carcinoma cell line,the expression of lnc RNAs was knocked out by si RNA transfection,observe the change of DCP level.Results:1.The expression levels of lnc RNA ATB、HULC、TMPO-AS1,CACNA1G-AS1、ZEB2-AS1 in serum of patients with low HBV-DNA viral load were significantly higher than those of patients with high HBV-DNA viral load(P <0.05).2.The expression levels of lnc RNA CACNA1G-AS1 and lnc RNA ATB in low DCP concentration group were significantly higher than those in high DCP concentration group(P <0.05).3.After transfection of si-CACNA1G-AS1,the concentration of DCP did not change significantly(P > 0.05),but after transfection of si-ATB,the concentration of DCP increased significantly(P<0.05).Conclusions:In this study,lnc RNA ATB down-regulated DCP expression in PLC/PRF/5 cells.It is suggested that lnc RNA ATB may be involved in the regulation of DCP expression in HBV-related HCC,which has potential value for further study of inhibiting the development of HCC with lnc RNA ATB as a target.