J4 ›› 2011, Vol. 37 ›› Issue (2): 255-259.
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FAN Li-mei,ZHANG Li-hui,CUI Song-hua,CUI Man-hua
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Abstract:
Abstract:Objective To investigate the effect of siRNA interfering glucose regulated protein78 (GRP 78) on cisplatin-resistance of human ovarian cancer SKOV3/ DDP cells,and clarify the biomechanism of the effect of GRP78 gene silencing on the cisplatin-resistance of human ovarian cancer SKOV3/DDP cells. Methods The pSilencerTM3.0-H1-GRP78 siRNA plasmid was constructed,then it was transfected into cancer cells by Lipofectamine 2 000. The expressions of GRP78 mRNA and protein were measured by RT-PCR and Western blotting. The expressions of caspase-4 protein and caspase-3 protein were detected by Western blotting. The apoptotic rate of cells was measured by flow cytometry. Results Compared with control group,the GRP78 gene and protein expression levels were markedly down-regulated following GRP78 siRNA plasmid transfection(P<0.05);Compared with untransfecting GRP78 siRNA plasmid group,the protein expressions of caspase-4 and caspase-3 in SKOV3/DDP cells in GRP78 siRNA plasmid transfection group induced by cisplatin treatment were sinificantly increased(P<0.05),and the apoptotic rate was also increased(P<0.05). Conclusion The suppression of GRP78 gene expression can increase the apoptotic rate of SKOV3/DDP cells through up-regulating the expressions of caspase-4 and caspase-3,so te suppression of GRP78 gene expression can decrease the drug resistance of SKOV3/DDP cells to cisplatin.
Key words: glucose regulated protein78;ovarian neoplasms;drug resistance;apoptosis
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