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.
ABSTRACT
In our previous study,we identified a novel testis-specific expressed gene 2(TSEG-2)from mouse testis.To further investigate its functions,35 male Balb/c mice(8 weeks old)were divided into cryptorchidism group(n=20),sham group(n=10),and control group(n=5).In cryptorchidism group,the right testes were anchored to the inner lateral abdominal wall.In situ hybridization(ISH)was applied to measure the localization of TSEG-2 in mouse testis.Real-time quantitative PCR was performed to detect the expression of TSEG-2 gene.Meanwhile,under the mediation of polyethylenimine(PEI),the recombinant vector pEGFP-TSEG-2(n=5)or empty vector(mock,n=5)was transfected into the testis of male mice.The transfection efficiencies were measured under a fluorescence microscope.The apoptosis of spermatogenic cells was detected by terminal deoxynuleotidyl-mediated nick end labeling(TUNEL).The results showed that TSEG-2 was expressed in convoluted seminiferous tubules,more precisely,in spermatogonia and spermatocytes.As compared with sham and control groups,the TSEG-2 transcription was significantly enhanced(P<0.05)and was correlated with apoptosis of spermatogenic cells in cryptorchid testes(P<0.05).PEI was efficient in mediating transfection of TSEG-2 into seminiferous tubules of testis.One week post-transfection,intratesticular injection of TSEG-2 resulted in increased apoptosis of spermatogenic cells in vivo (P<0.05).These results indicate that TSEG-2 may participate in the apoptosis of spermatogenic cells and the pathogenesis of cryptorchidism.
ABSTRACT
ysis, which had important reference values for further studying biological functions of UP Ⅱ gene and targeted therapeutic strategy for TCC of bladder.