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目的将靶向PKM2的siRNA转染食管癌细胞株Eca109,观察转染后细胞内PKM2基因的表达变化及其对Eca109细胞增殖和周期的影响。方法将人工合成的针对PKM2基因的siRNA片段通过Lipofectamine 2000转染食管癌细胞株Eca109;实验分为实验组、空白对照组和阴性对照组,分别用实时荧光定量聚合酶链反应(real-time quantita-tive PCR,qRT-PCR)和Western blot法检测各组细胞中PKM2的mRNA和蛋白质的表达量,并用MTT法检测各组细胞的增殖活力,流式细胞仪检测细胞周期的变化。结果与空白对照、阴性对照比较,实验组细胞中PKM2 mRNA及蛋白表达减少(P<0.05),Eca109细胞转染PKM2 siRNA后,增殖能力减弱,细胞周期被阻滞在G0/G1期。结论 PKM2对Eca109细胞的生长起促进作用。
Objective To transfect siRNA targeting PKM2 into esophageal carcinoma cell line Eca109 and observe the expression of PKM2 gene in transfected cells and its effect on the proliferation and cycle of Eca109 cells. Methods The synthetic siRNA targeting PKM2 gene was transfected into Eca109 cells by Lipofectamine 2000. The experiment was divided into experimental group, blank control group and negative control group. Real-time quantitative PCR The expression of PKM2 mRNA and protein in each group was detected by Western blot and qRT-PCR. The cell viability was measured by MTT assay. The cell cycle was detected by flow cytometry. Results Compared with the blank control and the negative control, the expression of PKM2 mRNA and protein in the experimental group was decreased (P <0.05). The proliferation of Eca109 cells was decreased and the cell cycle was arrested in the G0 / G1 phase. Conclusion PKM2 can promote the growth of Eca109 cells.