Smac作用于TRAIL诱导的前列腺癌PC-3细胞凋亡的研究.docVIP

  Effect of Smac on TRAIL-induced Apoptosis of Prostate Cancer Cell Line PC-3 and the Molecular Mechanism#  5 10 15 20 25 30 Wang Miao, Huang Tao, Wang Liang, Zeng Fuqing, Zheng Liduan, Tong Qiangsong** (Department of Urology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, WuHan 430022) Abstract: The effect of Smac gene on the TRAIL-induced apoptosis of the prostate cancer cell line PC-3 and the molecular mechanism were investigated. The Smac gene was transfected into PC-3 cells under the induction of liposome. The intrinsic Smac gene expression was detected by Western blot. After treatment with TRAIL as an apoptosis inducer, in vitro cell growth activity was assayed by MTT colorimetry. The apoptosis rate of PC-3 cells was determined by annexin V-FITC and propidium iodide staining flow cytometry. The expression of cellular XIAP and caspase-3 genes was examined by Western blot. Smac-transfected cells (PC-3/Smac group) had significantly increased Smac protein level as compared with PC-3 controls (P0.01). After induction with 100-200 ng/mL TRAIL for 12-36 h, cellular proliferation rate in PC-3/Smac group was significantly lower than in PC-3 controls (P0.05). After induction with 100 ng/mL TRAIL for 24 h, the apoptosis rate in PC-3/Smac group was significantly enhanced as compared with that of PC-3 controls (P0.05). Accordingly, the XIAP expression level was down-regulated significantly (P0.05) and caspase-3 subunit P20 was up-regulated significantly (P0.05). It is suggested that the overexpression of cellular Smac can inhibit inhibitor of apoptosis proteins (IAPs) , enhance caspases activity and the apoptosis rate of PC-3 cells induced by TRAIL, which may provide an useful experimental evidence for prostate cancer therapy. Keywords: Smac gene; prostate carcinoma; TRAIL; apoptosis Smac (the second mitochondria derived activator of caspase) or DIABLO (direct IAP binding protein with low pl), a mitochondrial protein that is rel