miR-296-5p靶向PLK1调控PI3K/AKT通路诱导骨肉瘤细胞中的自噬并抑制上皮-间质转化  

miR-296-5p Induces Autophagy in Osteosarcoma Cells and Inhibits Epithelial-mesenchymal Transition through Regulation of PI3K/AKT Pathways by Targeting PLK1

作　　者：孟爱霞[1] 官秀梅[1] 李桂芝[1] 孙风祥[1] MENG Aixia;GUAN Xiumei;LI Guizhi;SUN Fengxiang(Laboratory of Biochemistry and Molecular Biology,Weifang Medical University,Weifang,Shandong,261053,China)

机构地区：[1]潍坊医学院生物化学与分子生物学实验室,山东省潍坊市261053

出　　处：《医学分子生物学杂志》2022年第4期287-294,共8页Journal of Medical Molecular Biology

摘　　要：目的 探讨miR-296-5p靶向PLK1对骨肉瘤(osteosarcoma,OS)细胞自噬及抑制上皮-间质转化(EMT)的作用机制.方法 qRT-PCR检测miR-296-5p在OS细胞中的表达.采用生物信息学分析预测miR-296-5p的靶基因,验证miR-296-5p对靶基因PLK1的直接靶向调控;细胞转染构建miR-296-5p过表达和干扰细胞,CCK-8、克隆形成、Transwell小室、流式、蛋白免疫印迹实验检测miR-296-5p的不同表达对U2OS细胞中PTBP1表达水平及细胞增殖、侵袭、凋亡、自噬及EMT的影响.结果 与对照组比较,miR-296-5p在OS中表达降低,而PLK1则升高(P<0.05);与miR-NC组比较,mimic组的克隆形成率、侵袭细胞数目及PTBP1、p62、N-cadherin、Vimentin、p-PI3K/PI3K、p-AKT/AKT水平降低,细胞凋亡率、Bec-lin-1、LC3-Ⅱ/Ⅰ、E-cadherin水平升高(P<0.05);与PLK1组比较,PLK1+mimic组的克隆形成率、侵袭细胞数目及PTBP1、p62、N-cadherin、Vimentin、p-PI3K/PI3K、p-AKT/AKT水平降低,细胞凋亡率、Bec-lin-1、LC3-Ⅱ/Ⅰ、E-cadherin水平升高(P<0.05).结论 miR-296-5p可能能够靶向PLK1调控PI3K/AKT通路诱导OS细胞中的自噬并抑制EMT.Objective To explore the mechanism of miR-296-5p induced autophagy and inhibition of epithelial-mesenchymal transition(EMT)in osteosarcoma(OS)cells.Methods The expression level of miR-296-5p in OS cells was detected by qRT-PCR.The target gene of miR-296-5p was predicted by bioinformatic method.The direct binding relationship of miR-296-5p with its target gene PLK1 was verified.Cell lines overexpressed or silenced of miR-296-5p were constructed through cell transfection method.The effect of miR-296-5p on cell proliferation,invasion,apoptosis,autophagy,EMT,and the expression level of PTBP1 in U20S cells were detected by CCK-8,colony formation assay,transwell chamber assay,flow cytometry,and Western blotting.Results The expression level of miR-296-5p was decreased in OS,while the expression level of PLK1 was increased when compared to the control group(P<0.05).The colony formation rate,the number of invasive cells,and the expression levels of PTBP1,p62,N-cadherin,Vimentin,p-PI3K/PI3K and p-AKT/AKT were decreased in mimic group,while the apoptosis rate,the expression levels of Beclin-1,LC3-Ⅱ/Ⅰ and E-cadherin were increased when compared to the miR-NC group(P<0.05).The colony formation rate,the number of invasive cells,the expression levels of PT-BP1,p62,N-cadherin,Vimentin,p-PI3K/PI3K and p-AKT/AKT were decreased in PLK1+mimic group,while the apoptosis rate,the expression levels of Beclin-1,LC3-Ⅱ/Ⅰ and E-cadherin were increased in PLK1+mimic group when compared to the PLK1 group(P<0.05).Con clusion miR-296-5p may induce autophagy PI3K/AKT pathways by targeting PLK1.

关 键 词：骨肉瘤 miR-296-5p PLK1 PI3K/AKT通路 细胞自噬 上皮-间质转化 

分 类 号：R738.1[医药卫生—肿瘤]