Inhibition of hTERT gene by RNA interference in hepatoma cell line QGY

Objective To elucidate the time-efficiency relation of stable inhibition of hepatoma cells proliferation by RNA interference in vitro.Methods The stable screening-inhibition technique of RNAi was adopted to suppress the expression of hTERT stably.After small hairpin interfering RNA(shRNA) targeting hTERT gene was designed,recombinant vector pGenesil-shRNA-hTERT were constructed,and transfected into hepatoma QGY cells which were stably selected by G418 to establish hepatoma cell lines of stable expression pGenesil-shRNA-hTERT.Real-time RT-PCR,MTT and PCR-TRAP were utilized to detect the alterations of hTERT mRNA expressions,telomerase activity and cell proliferation.Results The effectiveness of RNAi existed continually and stably in hepatoma QGY cells expressing stably pGenesil-shRNA-hTERT.In the cell lines expressing stably pGenesil-shRNA-hTERT,hTERT mRNA was obviously suppressed,the telomerase activities were significantly decreased.Hepatoma QGY cell proliferation significantly inhibited in pGenesil-shRNA-hTERT groups compared to negative control groups.Conclusion RNAi may continually and stably suppress hTERT mRNA expressions and neoplasm proliferation,which is a potential new approach for neoplasm gene therapy.