【摘要】 通过设计并化学合成人端粒酶催化亚单位(hTERT)特异性siRNA,观察其对hTERT表达水平及肿瘤细胞生长的影响。将hTERT-siRNA以脂质体法转染入HeLa细胞,应用RT-PCR、实时定量TRAP、Western印迹、软琼脂克隆形成实验、荷瘤裸鼠肿瘤内注射等方法检测细胞内hTERTmRNA、蛋白质表达水平及对肿瘤细胞生长的影响。RT-PCR、实时定量TRAP和Western印迹的结果显示hTERT-siRNA明显降低了HeLa细胞内hTERT的mRNA及蛋白质表达水平并伴随有端粒酶活性的下降。克隆形成实验表明hTERT-siRNA组的体外肿瘤形成能力受到抑制。荷瘤裸鼠肿瘤内注射hTERT-siRNA使肿瘤平均体积显著小于对照组。TUNEL凋亡检测表明hTERT-siRNA转染组的凋亡率明显高于对照组。研究表明hTERT特异性siRNA可以明显抑制HeLa细胞内hTERT的表达水平,对其生长有明显抑制作用,是一种有前途的肿瘤治疗新方法。更多还原

【Abstract】 In this study, we aimed to observe the effect of chemically synthesized human telomerase catalytic subunit (hTERT)-oriented siRNA in cultured cell line and in a mouse xenograft model. Transfected hTERT- siRNA into HeLa cells, then using the RT-PCR, RQ TRAP, Western blot, soft agar colony forming assay, and intratumoral injection of hTERT-siRNA in a mouse xenograft model to investigate the level of hTERT mRNA and protein, the activity of telomerase, energy for growth of HeLa in vitro and in vivo. The result of RT-PCR, Western blot and RQ TRAP had showed that after transfected with hTERT-siRNA, the level of hTERT mRNA and protein decreased, and the activity of the telomerase was depressed obviously. The colony forming assay showed that energy for growth of hTERT-siRNA group was lower than that of the two control groups. The animal experiments showed that the cancer growth of hTERT-siRNA group was depressed. TUNEL assay showed an increased apoptosis rate in hTERT-siRNA group. Our study have showed that hTERT-siRNA can inhibit the expression of hTERT and depress the growth of the HeLa cells by inducing apoptosis in vitro and in vivo.更多还原