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OBJECTIVE:To investigate inhibitory effects of matrine on the proliferation of human bladder cancer BIU-87 cells and its mechanism. METHODS:The cell viability was detected by MTT assay and inhibitory rate of cell proliferation was calculat-ed after human bladder cancer BIU-87 cells were treated with 0(negative control),0.5,1.0,2.0 and 4.0 mg/ml matrine for 24,48 and 72 h,respectively. After treated with 0 (negative control),0.5,1.0,2.0 and 4.0 mg/ml matrine for 48 h,the cell cycle and apoptotic rate were detected by flow cytometry;the expression of Survivin,Caspase-3 and Caspase-7 protein were detected by Western blot assay. RESULTS:Compared with negative control,the proliferation of BIU-87 cells were significantly inhibited after incubated with 1.0-4.0 mg/ml matrine for 24,48 and 72 h(P<0.05 or P<0.01),and inhibitory rate of cell proliferation increased in concentration and time-dependant manner;after treated for 48 h,the percentage of G0/G1 phase cells and apoptotic rate in-creased,while the percentage of cells at S phase and G2/M phase were decreased (P<0.05 or P<0.01);the expression of Cas-pase-3 and Caspase-7 protein increased,while the expression of survivin protein decreased after incubated with 0.5-4.0 mg/ml ma-trine for 48 h. CONCLUSIONS:Matrine can inhibit the proliferation of human bladder cancer BIU-87 cells,block the cell cycle and induce apoptosis;its mechanism may be related to the expression regulation of Survivin,Caspase-3 and Caspase-7.
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Objective To explore the influence on the expression of elF4E and heparanase by antisense oligodeoxyribonucleotides (ASODN) transfection in human bladder carcinoma BIU-87 cells.Methods After transfecting the 2.5,5.O,7.5 μg/ml eIF4E ASODN into BIU-87 cells,at 24 h,48 h and 72 h,a cell control group (no transfection),a blank control group (empty liposomes) and a non-sense control group (transfected with non-sense ASODN) were established.The expression of eIF4E,HPA and mRNA were detected by in situ hybridration.The expression of eIF4 and HPA protein were detected by immunocytochemistry.SPSS 13.0 statistical software was used for statistical analysis.Results The expression quantity of eIF4E protein and mRNA were lower in the experimental groups ( protein:2.5 μg/ml ASODN treated 24 h,48 h,72 h were:3.55 ±0.52,3.52 ±0.51,3.22.±0.44 respectively; 5.0 μg/ml group:3.43 ±0.47,3.41 ± 0.46,3.19 ± 0.41 respectively ; 7.5 μg/ml group:2.36 ± 039,2.33 ± 0.37,2.05 ± 0.30 respectively.mRNA:2.5 μg/ml treated 24 h,48 h,72 h were:3.19 ±0.41,3.13 ±0.39,2.90 ±0.38 respectively ; 5.0 μg/ml group:3.07 ± 0.39,2.94 ± 038,2.27 ± 0.37 respectively ; 7.5 μg/ml group:2.22 ± 037,2.06 ± 0.30,1.95 ± 0.29 respectively.All data were less than those in the control groups (P ＜0.05).The expression quantity of HPA protein and mRNA were lower in experimental groups (protein:2.5 μg/ml ASODN treated 24 h,48 h,72 h were:4.44 ±0.55,4.40 ±0.54,3.99 ±0.52 respectively; 5.0 μg/ml group:4.41 ±0.55,4.21 ±0.53,3.77 ±0.50 respectively; 7.5 μg/ml group:4.02 ±0.52,3.98 ±0.52,2.32 ±0.37 respectively.mRNA:2.5 μg/ml treated 24 h,48 h,72 h were:4.12 ±0.51,3.75 ± 0.50,3.63 ± 0.45 respectively ; 5.0 μg/ml group:4.00 ± 0.51,3.71 ± 0.50,3.54 ± 0.44respectively ; 7.5 μg/ml group:3.87 ± 0.52,3.61 ± 0.49,2.08 ± 0.30 respectively).All data were less than in control groups ( P ＜ 0.05 ).There was a positive correlation on expression of HPA protein and eIF4E protein by inhibitory effect of eIF-4E ASODN (protein r=9.23,mRNA r=9.59,P ＜0.01).Conclusion eIF-4E ASODN might be used to inhibit the expression of eIF-4E gene and HPA gene in bladder cancer BIU-87 cells.
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Objective: To investigate the expression and identification of CH50 polypeptide in bladder cancer cell line BIU-87 by gene transfection. Methods:The eukaryotic expressing vector pCH510 of polypeptide CH50 was introduced into BIU-87 cells by gene transfection Lipofectimine TM2000.The expressed product was identified by immunohistochemistry and Western blot method. The expression of the transfected gene was identified by RT-PCR. Results:The BIU-87 cells could produce CH50 polypeptide by transfection. Conclusion:When the vector of pCH510 was transfected into BIU-87 cells in vitro, the cell adhension characteristic would be changed.