An Effect And Mechanism Study Of GTF3A On The Proliferation Of Hepatocellular Carcinoma Cells

Objective:The incidence of hepatocellular carcinoma(HCC)ranks the sixth among all cancers,and its mortality rate ranks the third,but its mechanism is not clear.GTF3A(General transcription factor,GTF3 A,or TFIIIA)is a general transcription factor of RNA polymerase III.The general transcription factors of RNA polymerase III includes TFIIIA?TFIIIB and TFIIIC.The role of TFIIIB and TFIIIC in the occurrence and development of cancer has been reported,but there are few studies on GTF3 A.This paper aims to explore the functions and molecular mechanisms of GTF3 A in the hepatocellular carcinoma,and provides a new molecular markers and targets for the diagnosis and treatment of HCC.Methods:(1)The HCC cell lines(Hep3B?Bel-7402)are stably knockdown by two specific shRNAs(short hairpin,RNA),and GTF3A-knockdown effects were measured by western-blotting and RT-PCR.(2)The cell proliferation capacity and colony ability of GTF3A-knockdown cell lines(shRNA-Hep3B?shRNA-Bel-7402)were examined by the MTT assay and the colony formation.(3)GTF3A-knockdown cell lines,shRNA-Hep3 B and shRNA-Bel-7402,were stained by PI(propidiumiodide,PI).And then,cell cycle profiles of the stained cell lines were measured by the FACS cytometry.CDK2 and cyclinD1 expressions in the GTF3A-knockdown cell lines were determined by western blotting.(4)The GTF3A-knockdown cell lines were stained by AnnexinV-FITC-PI.And then the apoptotic cell profiles were measured by FACS cytometry.Bcl-2 and cleaved caspase-3 in the GTF3A-knockdown cell lines were determined by western blotting.Results:(1)Western-blotting results showed that HCC cell lines,Hep3 B and Bel-7402 cell lines are stably knockdown by two specific shRNAs,and the protein and mRNAof GTF3 A are decreasing in shRNA-Hep3 B and shRNA-Bel-7402 cells when compared with the blank control.GTF3A-knockdown cell lines,shRNA-Hep3 B and shRNA-Bel-7402 were obtained and used in the next experiments.(2)The MTT assay and colony formation data showed that GTF3 A knockdown inhabits cell growth and colony formation in the HCC cell lines.(3)The cell cycle analysis showed that GTF3 A knockdown delays G1/S transition to arrest cell cycle of the HCC cell lines.The proteins expression levels of CDK2 and cyclinD1 are increased.(4)The apoptosis analysis showed that GTF3 A knockdown increases the apoptotic cells of the HCC cell lines Hep3 B.The proteins expression level of Bcl-2 is decreased,and cleaved caspase-3 is increased.Conclusions:GTF3A-knockdown inhibits the growth of HCC,and blocks HCC cell cycle.GTF3 A knockdown increases HCC cell apoptosis.GTF3 A may be an oncogene that promotes the development of HCC.GTF3 A may be as a potential molecular target for the treatment of HCC.