Modification of alternative splicing of Bcl-x pre-mRNA in bladder cancer cells
Zhu Zhaohui , Xing Shian , Cheng Ping , Zeng Fuqing , Lu Gongcheng
Current Medical Science ›› 2006, Vol. 26 ›› Issue (2) : 213 -216.
Modification of alternative splicing of Bcl-x pre-mRNA in bladder cancer cells
To modify the splicing pattern of Bcl-x and compare the effect of this approch with that of the antisense gene therapy in BIU-87 cell line of bladder cancer, by using 5′-Bcl-x AS to target downstream alternative 5′-Bcl-x splice site to shift splicing from Bcl-xL to Bcl-xS and 3′-Bcl-x AS AN tisense to the 3′-splice site of exon III in Bcl-x pre-mRNA to down regulation of Bcl-xL expression, the inhibitory effects on cancer cells by modification of alternative splicing and antisense gene therapy were observed and compared by microscopy, MTT Assay, RT-PCR, FACS, Westhern bloting and clone formation. The growth of cells BIU-87 was inhibited in a dose-and time-dependent maner. Its inhibitory effect began 12 h after the exposure, reaching a maximum value after 72 h. The number of cells decreased in S phase and the number increased in G1 phase. The ability of form foci was reduced and the antisense gene therapy was approximately half as efficient as modification of alternative splicing in inducing apoptosis. It is concluded that modification of splicing pattern of Bcl-x pre-mRNA in bladder canner cell BIU-87 is better than antisense gene therapy in terms of tumor inhibition.
bladder cancer / Bcl-x / alternative splicing
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