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目的:探讨PIK3CA在食管鳞癌细胞系Eca109中的作用.方法:用脂质体介导siRNA瞬时转染Ecal09细胞,转染细胞分为3组:空白对照组(正常培养的Eca109细胞)、阴性对照组(转染随机序列的siRNA)及实验组(转染PIK3CA-siRNA).运用Western blot技术检测PIK3CA基因干扰后蛋白水平表达的变化;四甲基偶氮唑盐微量酶反应比色法(MTT法)和细胞划痕实验检测PIK3CA干扰后Eca109细胞增殖和迁移能力的改变;流式细胞仪检测PIK3CA干扰后细胞周期和凋亡率的变化.结果:干扰PIK3CA基因表达后,其蛋白水平表达较空白对照组及阴性对照组显著性降低(P<0.05).MTT实验和划痕实验显示干扰PIK3CA的表达后,Eca109细胞增殖受到显著性抑制(P<0.05),迁移能力显著性降低(P<0.05).流式细胞仪检测下调P I K3C A的表达后细胞周期阻滞在细胞分裂的S期(P<0.05),且使细胞凋亡率显著性增加(P<0.05).结论:PIK3CA基因能够促进Eca109细胞的增殖和迁移能力,并具有抗凋亡作用.PIK3CA基因有望成为食管鳞癌治疗的潜在靶点.
Objective: To investigate the role of PIK3CA in esophageal squamous cell carcinoma cell line Eca109. METHODS: Transfect Ecal09 cells transiently with liposome-mediated siRNA. Transfected cells were divided into 3 groups: blank control group (normally cultured Eca109 cells), negative Control group (transfection of random sequence siRNA) and experimental group (transfection of PIK3CA-siRNA). Western blot technique was used to detect the change of protein level expression after interference of PIK3CA gene; tetramethylazolium salt trace enzyme reaction colorimetry ( MTT assay and cell scratch assay were used to detect the change of proliferation and migration ability of Eca109 cells after PIK3CA interference; Flow cytometry was used to detect the changes of cell cycle and apoptosis rate after PIK3CA interference. Results: After the PIK3CA gene expression was interfered, its protein level was expressed. Compared with the blank control group and the negative control group, there was a significant decrease (P<0.05). MTT assay and scratch test showed that after the interference of PIK3CA expression, the proliferation of Eca109 cells was significantly inhibited (P<0.05), and the migration ability was significantly decreased (P<0.05). <0.05) Flow cytometry detected PI K3C A downregulation of the cell cycle arrest in cell division S phase (P <0.05), and significantly increased the rate of apoptosis (P <0.05). Conclusion: PIK3CA Genes Promote the Proliferation of Eca109 Cells Migration, and has anti-apoptotic effects .PIK3CA is expected to be a potential target for gene therapy of esophageal squamous cell carcinoma.