论文部分内容阅读
目的:探讨抑制MicroRNA 21(miR-21)的表达对胃癌细胞的生物学行为及PTEN表达的影响,并分析miR-21参与肿瘤发生、发展的可能机制。方法:应用细胞瞬转的方法将miR-21 inhibitor转染课题组前期试验证实的miR-21高表达胃癌细胞株,应用细胞增殖试验(CCK8方法)、流式细胞技术(Annexin-Ⅴ标记)、Transwell试验检测抑制胃癌细胞株中miR-21表达后,对其细胞增殖、凋亡、周期、迁移的影响;并用Western印迹技术和双荧光素酶报告载体技术检测下调miR-21后,胃癌细胞株PTEN表达的变化。结果:体外增殖试验显示,下调miR-21后对胃癌细胞株的增殖抑制作用明显(P<0.05);体外凋亡试验显示,下调miR-21能增加胃癌细胞株的凋亡(P<0.05);在整个周期中以G1/S期为主;Transwell试验证实miR-21下调后,胃癌细胞株迁移能力明显下降(P<0.05);Western印迹试验显示,抑制miR-21表达后胃癌细胞株PTEN蛋白表达明显增加,荧光素酶相对活性明显增加(P<0.05)。结论:下调miR-21的表达对胃癌细胞株的增殖有明显的抑制作用,且促进其凋亡,能降低其体外迁移能力;下调miR-21后,PTEN活性明显增加;PTEN可能是miR-21参与胃癌发生、发展的靶标之一。
OBJECTIVE: To investigate the effect of miR-21 on the biological behavior and the expression of PTEN in gastric cancer cells. The possible mechanism of miR-21 in tumorigenesis and development was analyzed. Methods: miR-21 inhibitor was transfected into the gastric cancer cell lines with miR-21 overexpression confirmed by previous experiments in cell transfection. Cell proliferation assay (CCK8), Annexin-V labeling Transwell assay was used to detect the effect of miR-21 on the proliferation, apoptosis, cycle and migration of gastric cancer cells. Western blotting and dual luciferase reporter assay were used to detect the downregulation of miR-21. Changes in PTEN expression. Results: In vitro proliferation assay showed that downregulation of miR-21 inhibited the proliferation of gastric cancer cell lines significantly (P <0.05). In vitro apoptosis assay showed that downregulation of miR-21 can increase the apoptosis of gastric cancer cell lines (P <0.05) ; G1 / S phase was the main part of the whole cycle; Transwell assay confirmed that miR-21 down-regulated the migration ability of gastric cancer cell lines significantly decreased (P <0.05); Western blotting showed that inhibition of miR-21 expression in gastric cancer cell lines PTEN The protein expression was significantly increased, and the relative luciferase activity was significantly increased (P <0.05). Conclusion: The down-regulation of miR-21 expression significantly inhibited the proliferation of gastric cancer cell lines and promoted its apoptosis in vitro migration ability; down-regulation of miR-21, PTEN activity was significantly increased; PTEN may be miR-21 Involved in the occurrence and development of gastric cancer.