miR-96-5p靶向PTEN激活PI3K-Akt信号通路抑制非小细胞肺癌细胞增殖、迁移和侵袭实验研究

miR-96-5p targeting PTEN promotes non-small cell lung cancer migration,invasion,and proliferation by activating PI3K-Akt pathway

目的:探讨miR-96-5p对非小细胞肺癌(NSCLC)细胞迁移、侵袭和增殖的作用及分子机制。方法:在人NSCLC细胞系A549中转染miR-96-5p类似物(miR-96-5p mimics)、阴性对照(NC mimics)或miR-96-5p抑制剂(miR-96-5p inhibitor)、阴性对照(NC inhibitor),通过CCK-8法检测细胞的增殖能力,通过划痕实验和Transwell迁移实验检测细胞的迁移侵袭能力;采用荧光素酶报告基因实验检测miR-96-5p与磷酸酯酶与张力蛋白同源物(PTEN)启动子区DNA的结合;采用蛋白质免疫印迹实验检测miR-96-5p对PTEN/PI3K/Akt信号通路的调控作用。结果:抑制miR-96-5p, NSCLC细胞的迁移、侵袭和增殖能力降低,过表达miR-96-5p促进NSCLC细胞的迁移、侵袭和增殖(均P<0.05)。荧光素酶报告基因实验结果显示,miR-96-5p能与PTEN启动子区DNA结合;蛋白质免疫印迹实验结果表明,抑制miR-96-5p, PTEN表达升高、p-Akt的表达降低,过表达miR-96-5p后,PTEN表达降低、p-Akt的表达升高(均P<0.05),且沉默PTEN的表达后,逆转了敲低miR-96-5p对p-Akt的抑制作用(均P<0.05)。结论:miR-96-5p通过抑制PTEN,激活PI3K-Akt信号通路,促进NSCLC细胞的迁移、侵袭和增殖。

Objective:To investigate the effect and molecular mechanism of miR-96-5p on the migration,invasion and proliferation of non-small cell lung cancer(NSCLC)cells.Methods:Human NSCLC cell lin e A549 was transfected with miR-96-5p mimic(miR-96-5p mimics),negative control(NC mimics)or miR-96-5p inhibitor(miR-96-5p inhibitor),negative control(NC inhibitor).The proliferation ability of the cells was detected by CCK-8 method.The migration and invasion ability of the cells were detected by scratch test and Transwell migration test.Luciferase reporter assay was used to detect the binding of miR-96-5p to the promoter region of phosphatase and tensin homolog(PTEN)DNA.Western blot was used to detect the regulatory effect of miR-96-5p on PTEN/PI3K/Akt signaling pathway.Results:Inhibition of miR-96-5p reduced the migration,invasion and proliferation of NSCLC cells,while overexpression of miR-96-5p promoted the migration,invasion and proliferation of NSCLC cells(all P<0.05).Luciferase reporter gene assay showed that miR-96-5p could bind to the DNA of PTEN promoter region.Western blot results showed that inhibition of miR-96-5p increased the expression of PTEN and decreased the expression of p-Akt.After overexpression of miR-96-5p,the expression of PTEN decreased and the expression of p-Akt increased(all P<0.05).The inhibition effect of knockdown of miR-96-5p on p-Akt was reversed(P<0.05).Conclusion:miR-96-5p promotes the migration,invasion and proliferation of NSCLC cells by inhibiting PTEN and activating PI3K-Akt signaling pathway.