靶向生存素基因的shRNA抑制膀胱癌细胞裸鼠移植瘤生长

shRNA targeting survivin gene suppresses growth of bladder cancer cell xenografts in nude mice

目的观察稳定转染靶向生存素（survivin）基因短发夹RNA（shRNA）对膀胱癌细胞裸鼠移植瘤生长的影响。方法将靶向生存素基因的shRNA真核表达载体转染人膀胱癌细胞T24并获得稳定的单克隆细胞,实时定量PCR检测转染后生存素mRNA表达的变化;分别将转染和未转染细胞种植于裸鼠皮下,观察肿瘤生长情况;通过免疫组化检测肿瘤组织生存素蛋白表达的差异。结果RNA干扰使T24细胞生存素表达下调92.86%;未转染组在20～29d均形成体积超过2000mm3的肿瘤,转染组在43d时经病理证实3只裸鼠形成肿瘤,最大体积62.5mm3;免疫组化显示转染组形成的肿瘤生存素蛋白表达明显减弱。结论靶向生存素基因的shRNA明显下调了生存素表达,并抑制了膀胱癌细胞T24裸鼠移植瘤的生长。

Objective To study the effect of stable transfection of shRNA targeting survivin gene on the growth of bladder cancer cell T24 xenografts in nude mice. Methods Bladder cancer cell line T24 was transfected with survivin-targeting shRNA eukaryotic expression vector, and stable monoclonal ceils were selected. Survivin mRNA expression of transfeted and nontransfected ceils were detected by real time PCR. Transfected and nontransfected ceils were respectively inoculated into subcutaneous tissue of nude mice, and tumor growth was observed. Survivin protein expression of tumors was investigated by immunohistochemistry. Results Compared with nontransfected ceils, survivin mRNA expression of stably tansfected ceils was down-regulated by 92.86%. The tumor volume of the nontransfected group exceeded 2000 mm^3 between 20 and 29 days after inoculation, but the transfected group had only three tumors no more than 62.5 mm^3 after additional 2 weeks. Survivin protein expression of the tumors in transfected group was weaker than that in untransfected group. Conclusion Survivin-targeting shRNA markedly suppressed the growth of bladder cancer cell xenografts in nude mice through lastingly inhibition effect on survivin.