Effect Of Down-Regulation Of NUF-2 Gene Expression Mediated By RNA Interference On Proliferation And Cell Cycle Of Hepatocellular Carcinoma

Objective:Investigate the effect of down-regulation of NUF-2 gene expression on the proliferation and cell cycle of hepatocellular carcinoma(HCC).Materials and methods:HepG2?BEL-7404?SMMC-7721?BEL-7402 hepatocellular carcinoma cell lines and normal hepatocyte HL-7702 were selected for the study.The expression of HepG2,BEL-7404,SMMC-7721 and BEL-7402 in hepatocellular carcinoma cells and hepatocellular carcinoma tissues was detected by real-time fluorescence quantitative PCR(qRT-PCR).Then we selected the highly expressed hepatocellular carcinoma cell lines and made a construction mode of a lentivirus vector to repress the expression of NUF-2 gene by RNA interference,and infected the cell lines to obtain the stably inhibited expression of NUF-2.After successful lentivirus transfection,the expression of NUF-2 gene was determined by qRT-PCR.The results of qRT-PCR showed that the inhibited NUF-2 gene was used to select the highly expressed hepatocellularcarcinoma cells.The expression of NUF-2 protein in cancer cell lines was detected by Western blotting.Finally,the effect of inhibiting NUF-2 gene transfection on cell cycle was further explored,and the cell cycle was detected by flow cytometry.Results: The NUF-2 gene in four human hepatocellular carcinoma cell lines,expression:The real-time fluorescence quantitative PCR(qRT-PCR)results showed that the NUF-2 in four hepatocellular carcinoma cell lines,expression was significantly higher than that in normal hepatocellular carcinoma cell line HL-7702,of which SMMC-7721 was the most abundant.2.By inhibiting NUF-2 in SMMC-7721 human hepatocellular carcinoma cells mediated by lentiviruses,we found that the loss of NUF-2 in vitro significantly inhibited the ability of hepatocellular carcinoma protein formation.In addition,inhibition of NUF-2 can induce cell cycle arrest and increase of hepatocellular carcinoma cells in G1 phase.Conclusion : The expression of NUF-2 gene was remarkablely enhanced in human hepatocellular carcinoma,The ability of proliferation and invasion in SMMC-7721 cells can be inhibited by down-regulation of NUF-2,and increase the number of hepatocellular carcinoma in G1 phase.