Department of Urology, Seoul National University Hospital, Seoul, Korea.
BJU Int. 2010 Dec;106(11):1812-6. doi: 10.1111/j.1464-410X.2010.09259.x.
To investigate the effects of survivin gene RNA interference on cell growth and the cell cycle in the human bladder cancer cell line T24.
A small interfering RNA (siRNA) targeting survivin was transfected into T24 cells using a liposome approach. Reverse transcription-polymerase chain reaction and Western blot analysis were used to examine survivin gene expression in T24 cells. Cells densities were determined by haematocytometer counts and flow cytometry was used for cell cycle analysis. Caspase-3 activity was quantified.
After treatment with survivin siRNA, the survivin gene expression in T24 cells was almost completely absent. The survivin siRNA treatment caused a profound decrease in survivin protein, which was correlated with a decrease in cell growth, G2/M arrest, and an increase in the fraction of cells undergoing apoptosis. The inhibition of survivin expression increased caspase-3 activity in T24 cells, which led to apoptosis.
RNA interference can efficiently suppress survivin expression in T24 cells. Targeting survivin by siRNA may be a promising approach to block proliferation of bladder cancer cells and may provide a suitable adjuvant therapy for treatment of bladder cancer.
研究 RNA 干扰 Survivin 基因对人膀胱癌 T24 细胞生长和细胞周期的影响。
采用脂质体法将靶向 Survivin 的小干扰 RNA(siRNA)转染至 T24 细胞。采用逆转录-聚合酶链反应和 Western blot 分析检测 T24 细胞中 Survivin 基因的表达。通过血球计数器计数测定细胞密度,采用流式细胞术分析细胞周期。定量检测半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)活性。
经 Survivin siRNA 处理后,T24 细胞中的 Survivin 基因表达几乎完全缺失。Survivin siRNA 处理导致 Survivin 蛋白明显减少,与细胞生长减少、G2/M 期阻滞以及凋亡细胞比例增加相关。抑制 Survivin 表达增加了 T24 细胞中的 Caspase-3 活性,导致细胞凋亡。
RNA 干扰可有效抑制 T24 细胞中 Survivin 的表达。通过 siRNA 靶向 Survivin 可能是阻止膀胱癌细胞增殖的一种很有前途的方法,并可为膀胱癌的治疗提供一种合适的辅助治疗手段。