摘要
目的观察靶向survivin的siRNA对膀胱癌细胞T-24增殖和凋亡的影响。方法使用lipofectamineTM2000将靶向survivin的siRNA真核表达载体转染至膀胱癌细胞T-24,半定量RT-PCR检测T-24细胞survivin基因表达的变化;MTT法检测对T-24细胞增殖的影响;膜联蛋白V-PI(annexinV-PI)染色及流式细胞技术检测诱导凋亡的影响。结果靶向survivin的序列特异性siRNA可以高效率抑制T-24细胞survivin基因在基因水平的表达,mRNA抑制率为61.73%;细胞重新接种24、48h后,其增殖抑制率分别为32.42%和37.48%;转染24h后可以诱导细胞凋亡16.76%。结论利用RNA干扰技术沉默survivin基因表达可以显著抑制T-24细胞增殖,并在一定程度上诱导其自发凋亡,靶向survivin的RNA干扰技术在膀胱癌的基因治疗中具有一定价值。
Objective To investigate the effect of blocking the survivin expression with RNA interference（RNAi）techniques on supperresing proliferation and inducing apoptosis of bladder cancer T-24 cells. Methods A siRNA directed against survivin was transfeted into bladder cancer T-24 cells with lipofectamineTM 2000. The changes of survivin expression were detected by semi-quantitive RT-PCR. The effect of suppressing proliferation of T-24 cell was detected by MTT assay. The effect of inducing T-24 cell apoptosis was detected by flow cytometry. Results The sequence-specific siRNA efficiently blocked the survivin expression at mRNA level. The expression inhibition rate was 61.73 % at mRNA level detected by semi-quantitive RT-PCR. To blocking the survivin expression could suppress proliferration of T-24 cells significantly. At 24h and 48h after the cells were reseeded, the proliferation inhibition rates were 32.42% and 37.48% ,and at 24h after transfection,apoptosis was induced in 16.76% of the cells as detected by flow cytometry assay. Conclusion Blocking the survivin expression with RNAi technology can significantly suppress proliferation of T-24 cells and induce apoptosis to a certain degree. RNAi targeted to survivin has a potential value in gene therapy of bladder cancer.
出处
《重庆医学》
CAS
CSCD
2007年第14期1360-1362,共3页
Chongqing medicine
基金
重庆市教委基金资助项目(040313)
作者简介
通讯作者
