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目的:探讨RNA干扰Med19的表达对结肠癌Caco-2细胞增殖和凋亡的影响。方法:构建靶向Med19的干扰质粒pSilencer-Med19-siRNA,转染Caco-2细胞后,RT-PCR检测Caco-2细胞中Med19 mRNA的表达,Western blotting检测Caco-2细胞中Med19蛋白的表达,MTT检测pSilencer-Med19-siRNA转染后Caco-2细胞的增殖,流式细胞术分析Caco-2细胞的凋亡。结果:RT-PCR及Western blotting检测结果显示,pSilencer-Med19-siRNA转染后Caco-2细胞中Med19 mRNA及蛋白表达水平上均显著下降(P<0.01);MTT及流式细胞术检测结果表明,与pSilencer对照组相比,pSilencer-Med19-siRNA组Caco-2细胞的增殖明显受到抑制[7 d时:(0.86±0.09)%vs(1.38±1.10)%,P<0.01],且细胞凋亡比例明显升高[(22.72±2.85)%vs(7.23±1.29)%,P<0.01]。结论:RNA干扰沉默Med19的表达可抑制结肠癌Caco-2细胞的增殖,促进其凋亡,提示Med19可作为结肠癌治疗的潜在靶点。
Objective: To investigate the effect of RNA interference Med19 expression on the proliferation and apoptosis of Caco-2 cells. Methods: The expression plasmid pSilencer-Med19-siRNA targeting Med19 was constructed and transfected into Caco-2 cells. The expression of Med19 mRNA in Caco-2 cells was detected by RT-PCR. The expression of Med19 protein in Caco-2 cells was detected by Western blotting. The proliferation of Caco-2 cells after transfection with pSilencer-Med19-siRNA was detected by MTT. The apoptosis of Caco-2 cells was analyzed by flow cytometry. Results: The results of RT-PCR and Western blotting showed that the expression of Med19 mRNA and protein in Caco-2 cells was significantly decreased after transfection with pSilencer-Med19-siRNA (P <0.01). The results of MTT assay and flow cytometry , The proliferation of Caco-2 cells was significantly inhibited in pSilencer-Med19-siRNA group compared with pSilencer control group [(0.86 ± 0.09)% vs (1.38 ± 1.10)%, P <0.01 at 7 d (22.72 ± 2.85)% vs (7.23 ± 1.29)% respectively, P <0.01]. Conclusion: The silencing of Med19 by RNA interference can inhibit the proliferation and promote the apoptosis of Caco-2 cells, suggesting that Med19 can be used as a potential target for the treatment of colon cancer.