Li Yan, Xiao Li-ying, Yao Gang, Li Hong, Yang Zhi-wei, Li Wan-yi, Jiang Zhong-hua, Li Ming-yuan
Sichuan University, Chengdu, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008 Jun;24(6):570-3.
To study the effect of the hTR-siRNA adenovirus on hTR mRNA gene silence, telomerase activity inhibition and anti-tumor in vitro.
RNAi adenovirus vector, Ad-hTR-siRNA, and negative control Ad-NT-siRNA were constructed by an improved ligation method. Different tumor cells and liver cell line, HL-7702, were infected with 100 MOI of the recombinant adenoviruses. TRAP-ELISA, Real-time PCR and FCM were used to analyze telomerase activity, hTR mRNA, apoptosis rate and hTERT protein expression.
As compared with Ad-NT-siRNA, Ad-hTR-siRNA reduced both hTR mRNA levels (70.21%) and telomerase activity (58.87%) of HeLa cells significantly, increased apoptosis rate (29.7%). But the telomerase activity of HL-7702 and hTERT protein didn't show the tendency of decrease.
It is supposed that the hTR-siRNA adenovirus could knockdown hTR gene specifically and suppress the tumor cell growth in vitro efficiently. Maybe this siRNA expressing recombinant adenovirus system could be a new method for cancer gene therapy.
研究hTR - siRNA腺病毒对hTR mRNA基因沉默、端粒酶活性抑制及体外抗肿瘤的作用。
采用改良连接法构建RNAi腺病毒载体Ad - hTR - siRNA和阴性对照Ad - NT - siRNA。用100 MOI的重组腺病毒感染不同肿瘤细胞及肝细胞系HL - 7702。采用端粒重复序列扩增法 - 酶联免疫吸附测定（TRAP - ELISA）、实时荧光定量聚合酶链反应（Real - time PCR）和流式细胞术（FCM）分析端粒酶活性、hTR mRNA、凋亡率及hTERT蛋白表达。
与Ad - NT - siRNA相比，Ad - hTR - siRNA显著降低了HeLa细胞的hTR mRNA水平（70.21%）和端粒酶活性（58.87%），增加了凋亡率（29.7%）。但HL - 7702的端粒酶活性及hTERT蛋白未呈现下降趋势。
推测hTR - siRNA腺病毒可特异性敲低hTR基因并有效抑制体外肿瘤细胞生长。该表达siRNA的重组腺病毒系统可能成为癌症基因治疗的新方法。
