Study On The Mechanism Of TRB3 Inhibiting Proliferation Of Human Hepatocellular Carcinoma Cell Line MHCC-97H

Objective:To explore the expression and function of TRB3 in human hepatocellular carcinoma cell(HCC)MHCC-97H,and study its possible mechanism of inhibiting proliferation of MHCC-97H.Methods:TRB3 expressions in normal hepatocellular cell line L-02 and HCC MHCC-97H were assayed by real-time PCR.The expressions of TRB3 and CHOP were knock down by lentivirus containing short hairpin RNA(shRNA).Cell counting kit-8(CCK-8)experiment was used for testing cell growth and death in HCC MHCC-97H in which TRB3 had been knocked down.The expression of TRB3 and its related factors in HCC MHCC-97H in which TRB3 or CHOP had been knocked down,such as CHOP and AKT,were analyzed with Western blot and real time PCR respectively.Results:TRB3 expression in HCC MHCC-97H increased 3.8 fold compared with normal hepatocellular cell line L-02(p<0.05),CCK-8 experiment showed that the cell growth was enhanced in HCC MHCC-97H of low expression of TRB3(p<0.05).After CHOP gene was suppressed,the expressions of TRB3 mRNA and protein decreased.Similarly,the expression of CHOP mRNA and protein was down-regulated after TRB3 gene was suppressed(p<0.05).While TRB3 was down-regulated in HCC MHCC-97H,AKT mRNA was reduced(p<0.05),and its protein decreased but did not have statistical significance(p>0.05).However,Western blot illustrated the obvious rising of pAKT(p<0.05).Conclusion:TRB3 gene was up-regulated in HCC MHCC-97H,and could modified the cell death.Moreover,the mechanism of HCC MHCC-97H growth regulated by TRB3 was related to interacting with endoplasmic reticulum stress(ERS)factor CHOP and inhibiting AKT phosphorylation.TRB3 has theoretical value in HCC diagnosis and therapy.