MiR-372通过抑制PI3K/AKT/mTOR信号通路调节786-O细胞的增殖、凋亡和迁移  

MiR-372 regulates the proliferation,apoptosis and migration of 786-O cell by inhibiting PI3K/AKT/mTOR signaling pathway

作　　者：王鑫 张玉 纪世琪 刘庆军 Wang Xin;Zhang Yu;Ji Shiqi;Liu Qingjun(Department of Urology,Beijing Ditan Hospital,Capital Medical University,Beijing 100102,China)

机构地区：[1]首都医科大学附属北京地坛医院泌尿外科,北京100102

出　　处：《国际外科学杂志》2024年第6期413-418,F0003,F0004,共8页International Journal of Surgery

基　　金：北京医学奖励基金(YXJL-2021-0800-0410)。

摘　　要：目的探讨miR-372在不同表达情况下是否可以影响肾透明细胞癌(ccRCC)细胞增殖、迁移以及凋亡,并研究miR-372对PI3K/AKT/mTOR信号通路的影响。方法选取ccRCC细胞系(786-O)以及正常肾上皮细胞系(HK-2),采用实时荧光定量聚合酶链反应(RT-qPCR)检测786-O细胞和HK-2细胞中miR-372的表达水平。通过转染模拟物制成模拟物组及阴性对照组,通过转染抑制剂制成抑制剂组及阴性对照组。得到稳定转录的细胞系后通过细胞划痕实验检测迁移情况,CCK8法检测细胞增殖情况,以及流式细胞术检测细胞凋亡情况,再通过Western blotting法检测细胞系中增殖和凋亡相关蛋白以及PI3K/AKT/mTOR信号通路相关蛋白的表达水平。计量资料以均数±标准差(x±s)表示,组间比较采用独立样本t检验。结果在786-O细胞中,miR-372的表达水平降低,miR-372过表达抑制了786-O细胞中增殖细胞核抗原的表达,而促进了Bcl-2及Caspase3的表达,并可以降低PI3K/AKT/mTOR信号通路的磷酸化水平。结论miR-372能够抑制786-O细胞的迁移和增殖,促进细胞凋亡。此外,miR-372还可以抑制PI3K/AKT/mTOR信号通路。Objective To explore whether miR-372 can affect the proliferation,migration and apoptosis of renal clear cell carcinoma(ccRCC)cell under different expression conditions,and to study the effect of miR-372 on PI3K/AKT/mTOR signaling pathway.Methods ccRCC cell line(786-O)and normal renal epithelial cell line(HK-2)were selected to detect the expression level of miR-372 in 786-O cell and HK-2 cell by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).The mimic group and negative control group were prepared by transfection with the mimic agent,and the inhibitor group and negative control group were prepared by transfection with the inhibitor.After stable transcription of cell lines was obtained,cell migration was detected by cell scratch assay,cell proliferation was detected by CCK8 method,and cell apoptosis was detected by flow cytometry,and the expression levels of proliferation and apoptosis related proteins and PI3K/AKT/mTOR signaling pathway-related proteins were detected by Western blotting.Measurement data were expressed as mean±standard deviation(x±s),and independent sample t-test was used for comparison between groups.Results The expression level of miR-372 was decreased in 786-O cell.Overexpression of miR-372 inhibited the expression of proliferating cell nuclear antigen,promoted the expression of Bcl-2 and Caspase3,and reduced the phosphorylation level of PI3K/AKT/mTOR signaling pathway in 786-O cell.Conclusions miR-372 can inhibit the migration and proliferation of 786-O cell,and promote cell apoptosis.In addition,miR-372 can inhibit the PI3K/AKT/mTOR signaling pathway.

关 键 词：微RNA 细胞增殖 细胞凋亡 细胞迁移分析 MiR-372 肾透明细胞癌 

分 类 号：R737.11[医药卫生—肿瘤]