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Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 625-630, 2009.
Article Dans Chinois | WPRIM | ID: wpr-341170

Résumé

The recombinant plasmids pGenesii-1-BIRC71 and pGenesil-1-BIRC72 were transfected into Hela cells and cisplatin was added with different concentrations in order to study the inhibitory ef-fects of Livin gene, increase the apoptosis induced by cisplatin, and detect the expression of Bcl-2, Bax,caspase-3, and survivin genes. The pGenesil-1-BIRC71 and pGenesil-1-BIRC72 were transfected into Hela cells, and the expression levels of Livin, Bcl-2, Bax, caspase-3, and survivin genes were detected by using fluorescence quantitative real-time PCR. Then cisplatin at different concentrations (3.0, 6.0 and 9.9 μg/mL) was added into the transfected Hela cells, and 24, and 48 h later, the apoptosis rate was measured by flow cytometry. After transfection of pGenesil-1-BIRC71 and pGenesil-1-BIRC72 into Hela cells, the expression level of Livin gene was obviously reduced, and the apoptosis rate was sig-nificantly increased in transfection group as compared with control group (P<0.05). Cisplatin could in-crease the apoptosis rate in a dose- and time-dependent manner. After cisplatin was added, the expres-sion levels of Bcl-2 mRNA were reduced, and those of Bax, caspase-3, and survivin mRNA were in-creased in transfection group as compared with those in control group (P<0.05). It was concluded that shRNA expression vector targeting Livin gene could inhibit the expression of Livin gene in Hela cells and enhance the apoptosis induced by cisplatin, which was related to the decreased expression of Bcl-2and activation of Bax and caspase-3. Survivin might play an important role as an antagonist in the proc-ess of apoptosis induction.

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