【摘要】 目的研究针对人类端粒酶催化亚单位(hTERT)调控区C-MYC结合位点的反义RNA抑制肝癌细胞生长。方法细菌内同源重组构建反义RNA腺病毒(rAd-asmycb),转染HepG2.2.15肝癌细胞,流式细胞术检测细胞凋亡,透射电镜下观察凋亡细胞形态,聚合酶链反应-酶联免疫分析(PCR-ELISA)、逆转录-PCR(RT-PCR)分别检测细胞端粒酶活性及mRNA水平上hTERT表达。结果反义RNA抑制肝癌细胞生长,细胞凋亡率为40.7%,显示特征性凋亡细胞形态,能够显著降低细胞端粒酶活性,在mRNA水平上抑制hTERT表达。结论hTERT调控区C-MYC结合位点可能是肝癌基因治疗的有效靶位。更多还原

【Abstract】 Objective To study the inhibition effects on hepatoma cells growth by the anti-sense RNA targeting C-MYC binding site on regulation region of hTERT promoter.Methods The rAd virus which express anti-sense RNA complementary to the C-MYC binding site on regulation region of hTERT were constructed using the method of homologous recombination in bacteria cells.The apoptosis of HepG2.2.15 cells infected by rAd-asmycb was detected by the method of Annexin V-FITC/PI labeling,and the morphological changes were observed by electronic microscopy.TRAP-PCR-ELISA and RT-PCR were used to detecte the relative telomerase activity(RTA) and gene transcription at mRNA level of hTERT.Results Cell growth of HepG2.2.15 was retarded and about 40.7% tumor cells were lead to apoptosis.RTA of anti-sense RNA treated cells(1.175) was much lower than the control cells(4.200,P<0.05).RT-PCR showed gene transcription of hTERT was inhibited.Conclusion The C-MYC binding site of hTERT gene may be a target for HCC gene therapy.更多还原