miR-28-5p通过靶向MTSS1调控非小细胞肺癌细胞的恶性生物学行为

miR-28-5p regulates the malignant biological behavior of non-small cell lung cancer cells by targeting MTSS1

[目的]探讨miR-28-5p通过靶向MTSS1调控非小细胞肺癌细胞的恶性生物学行为。[方法]用双萤光素酶检测试剂分析荧光素酶活性,同时应用RT-PCR检测MRC5细胞和A549细胞中miR-28-5p和MTSS1 mRNA水平;用Lipofectamine法将miR-NC、miR-28-5p inhibitor和miR-28-5p mimic转染到A549细胞,培养48h后分别采用MTT法、流式细胞术和Transwell法检测细胞增殖、凋亡和迁移情况,同时蛋白印迹法法检测A549细胞中MTSS1、PI3K、AKT和Caspase-3表达。[结果] miR-28-5p与MTSS1有潜在的结合位点;转染miR-28-5p mimic可明显降低MTSS1-WT的荧光素酶活性,对MTSS1-MUT没有影响,而miR-28-5p inhibitor则表现出相反作用,表明miR-28-5p与MTSS1存在靶向调节作用;A549细胞中miR-28-5p mRNA的相对表达量高于MRC5细胞(P<0.05),A549细胞中MTSS1 mRNA的相对表达量低于MRC5细胞(P<0.05);通过miR-28-5p inhibitor降低miR-28-5p后,A549细胞凋亡、MTSS1、MTSS1、PI3K、AKT和Caspase-3的表达增加,细胞增殖和迁移降低(P<0.05);通过miR-28-5p mimic增加miR-28-5p后,A549细胞凋亡、MTSS1、MTSS1、PI3K、AKT和Caspase-3的表达降低,细胞增殖和迁移增加(P<0.05)。[结论] miR-28-5p在A549细胞中高表达,通过基因干预降低miR-28-5p表达后,miR-28-5p通过靶向MTSS1的3’端非编码区,提高MTSS1的翻译水平,抑制A549细胞的恶性生物学行为。

[Objective]To investigate the effect of miR-28-5 p on regulating the malignant biological behavior of non-small cell lung cancer cells by targeting MTSS1.[Method]The luciferase activity was analyzed with dual luciferase detection reagent,and RT-PCR was used to detect the levels of miR-28-5 p and MTSS1 mRNA in MRC5 cells and A549 cells.miR-NC,miR-28-5 p inhibitors and miR-28-5 p mimic were transfected into A549 cells by Lipofectamine method.After 48 hours of culture,cell proliferation,apoptosis and migration were detected by MTT method,flow cytometry and transwell method respectively.At the same time,Western Blotting method was used to detect MTSS1,PI3 K,AKT and Caspase-3 expression in A549 cells.[Result]miR-28-5 p had a potential binding site with MTSS1.Transfection of miR-28-5 p mimic can significantly reduce the luciferase activity of MTSS1-WT,and had no effect on MTSS1-MUT,while miR-28-5 p inhibitor showed the opposite effect,which indicated that miR-28-5 p and MTSS1 had a targeted regulation effect.The relative expression of miR-28-5 p mRNA in A549 cells was higher than that in MRC5 cells(P<0.05),and the relative expression of MTSS1 mRNA in A549 cells lower than MRC5 cells(P<0.05).After miR-28-5 p reduced by miR-28-5 p inhibitor,A549 cell apoptosis,MTSS1,MTSS1,PI3 K,AKT and Caspase-3 expression were increased,and cell proliferation and migration were decreased(P<0.05).After miR-28-5 p increased by miR-28-5 p mimic,A549 cell apoptosis,MTSS1,MTSS1,PI3 K,AKT and Caspase-3 expression were decreased,and cell proliferation and migration were increased(P<0.05).[Conclusion]miR-28-5 p is highly expressed in A549 cells.After reducing the expression of miR-28-5 p through genetic intervention,miR-28-5 p increases the translation level of MTSS1 and inhibits the malignant biological behavior of A549 cells by targeting the 3′non-coding region of MTSS1.