SiRNA稳定抑制肝癌细胞hTERT基因的表达

【关键词】  肝肿瘤

    Stable inhibition of hTERT gene by siRNA in hepatocarcinoma cells

　　【Abstract】 AIM: To elucidate the timeefficiency relation of stable inhibition of hepatocarcinoma cell proliferation by RNA interference in vitro. METHODS:  The stable screeninginhibition technique of RNAi was adopted to suppress the expression of hTERT stably. After small hairpin   RNA (shRNA) targeting hTERT gene was designed, recombinant vector pGenesilshRNAhTERT was constructed and transfected into hepatocarcinoma SMMC7721 cells, which were stably selected by G418 to establish hepatocarcinoma cell lines  stablely  expressing  pGenesilshRNAhTERT. Realtime RTPCR, MTT and PCRTRAP 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 hepatocarcinoma SMMC7721 cells expressing stably pGenesilshRNAhTERT. In the cell lines expressing stably pGenesilshRNAhTERT, hTERT mRNA was obviously suppressed, and the telomerase activities were significantly decreased. Hepatocarcinoma SMMC7721 cell proliferation significantly inhibited in pGenesilshRNAhTERT group compared to that in negative control group. CONCLUSION:  RNAi may continually and stably suppress hTERT mRNA expression and carcinoma cell  proliferation, which is a potential new approach for antitumor  gene therapy.

　　【Keywords】 RNA interference; liver neoplasms; telomerase

　　【摘要】  目

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