Effect of ZEB2 silencing on cisplatin resistance in gastric cancer.
D-M Geng, Kan Xm, Zhang Ww
Abstract
D-M Geng, Kan Xm, Zhang Ww
Abstract
OBJECTIVE: To investigate the effect of ZEB2 silencing on cisplatin resistance in gastric cancer. MATERIALS AND METHODS: The resulting cell line, SGC7901/DDP, was transfected with ZEB2 siRNA, non-specific siRNA, or vehicle control. The effectiveness of ZEB2 silencing was evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot. MTT viability assay was used to determine the cisplatin-sensitivity of cells. Cell apoptosis was measured by flow cytometry. RESULTS: A significant decrease in ZEB2 in mRNA and protein level was seen in cells transfected with ZEB2 siRNA, compared to that in cells transfected with non-specific siRNA or vehicle. Transfection with ZEB2 siRNA in cisplatin-resistant SGC7901/DDP cells resulted in a significant decrease in cell viability in response to the cisplatin treatment, and cell viability decreased with increasing cisplatin concentrations. A higher apoptotic rate was also seen in cells transfected with ZEB2 siRNA under cisplatin treatment. CONCLUSIONS: ZEB2 silencing can effectively make gastric cells sensitive to cisplatin treatment in vitro.
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OBJECTIVE: To investigate the effect of ZEB2 silencing on cisplatin resistance in gastric cancer. MATERIALS AND METHODS: The resulting cell line, SGC7901/DDP, was transfected with ZEB2 siRNA, non-specific siRNA, or vehicle control. The effectiveness of ZEB2 silencing was evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot. MTT viability assay was used to determine the cisplatin-sensitivity of cells. Cell apoptosis was measured by flow cytometry. RESULTS: A significant decrease in ZEB2 in mRNA and protein level was seen in cells transfected with ZEB2 siRNA, compared to that in cells transfected with non-specific siRNA or vehicle. Transfection with ZEB2 siRNA in cisplatin-resistant SGC7901/DDP cells resulted in a significant decrease in cell viability in response to the cisplatin treatment, and cell viability decreased with increasing cisplatin concentrations. A higher apoptotic rate was also seen in cells transfected with ZEB2 siRNA under cisplatin treatment. CONCLUSIONS: ZEB2 silencing can effectively make gastric cells sensitive to cisplatin treatment in vitro.
Key concepts: Cisplatin, Transfection, Viability assay, Gene silencing, Apoptosis, MTT assay, Flow cytometry, Molecular biology