Effect of Smac on TRAIL-induced apoptosis of prostate cancer cell line PC-3 and the molecular mechanism |
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Authors: | Miao Wang Tao Huang Fuqing Zeng Guosong Jiang Liang Wang Liduan Zheng Qiangsong Tong |
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Institution: | Department of Urology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China. wangmiao0825@gmail.com |
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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 blotting. 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 blotting.
Smac-transfected cells (PC-3/Smac group) had significantly increased Smac protein level as compared with PC-3 controls (P<0.01). 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 (P<0.05). 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 (P<0.05). Accordingly, the XIAP expression level was down-regulated significantly (P<0.05) and caspase-3 subunit P20 was up-regulated significantly (P<0.05). It is suggested that the over-expression 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 a useful experimental basis for
prostate cancer therapy. |
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Keywords: | Smac gene prostate carcinoma TRAIL apoptosis |
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