miR-145通过mTOR途径调控NSCLC A549细胞生物学特性  

miR-145 regulates the biological characteristics of NSCLC A549 cells through mTOR pathway

作　　者：李柱[1] 吴烜[1] 彭小丹[1] 王树滨[1] LI Zhu;WU Xuan;PENG Xiaodan;WANG Shubin(Department of Oncology,Shenzhen Hospital,Peking University,Shenzhen,Guangdong 518000,China)

机构地区：[1]北京大学深圳医院肿瘤内科,广东省深圳市518000

出　　处：《中南医学科学杂志》2022年第2期179-183,共5页Medical Science Journal of Central South China

基　　金：深圳市科技计划项目(ZDSYS20190902092855097)。

摘　　要：目的探究miR-145调控非小细胞肺癌(NSCLC)A549细胞生物学特性及对雷帕霉素靶蛋白(mTOR)信号转导通路的影响。方法将A549细胞分为miR-145组、对照组、miR-145抗体组和对照抗体组。采用Lipofectamine;2000转染试剂盒进行质粒转染,行MTT、细胞凋亡、细胞迁移、细胞侵袭检测,Western blotting检测分析雷帕霉素靶蛋白(mTOR)、磷酸化mTOR(p-mTOR)、真核细胞翻译起始因子(eIF4E)、磷酸化eIF4E(p-eIF4E)、核糖体蛋白S6激酶(p70S6K)、磷酸化p70S6K(p-p70S6K)、S6核糖体蛋白(S6)、磷酸化S6(p-S6)、eIF4E结合蛋白1(4E-BP)、磷酸化4E-BP(p-4E-BP)水平。结果对照组和对照抗体组细胞miR-145表达和细胞凋亡水平差异无显著性(P>0.05),但与miR-145组和miR-145抗体组比较有明显差异,其中miR-145组最高,miR-145抗体组最低(P<0.05)。对照组和对照抗体组细胞增殖、迁移、侵袭活性,以及p-mTOR/mTOR、p-eIF4E/eIF4E、p-p70S6K/p70S6K、p-S6/S6、p-4E-BP/4E-BP表达水平差异无显著性(P>0.05),但与miR-145组和miR-145抗体组比较有明显差异,其中miR-145组最低,miR-145抗体组最高(P<0.05)。结论miR-145可通过mTOR途径调控NSCLC A549细胞生物学特性,促进细胞凋亡,抑制细胞增殖活性、迁移能力及侵袭能力。Aim To analyze the cell biological characteristics of miR-145 regulating non-small-cell lung cancer(NSCLC)A549 and its effect on mammalian target of rapamycin(mTOR)signal transduction pathway.Methods A549 cells were divided into miR-145 group,control group,miR-145+antagonists group and control+antagonists group.The cells were transfected with plasmid by Lipofectamine;2000 transfection kit,and were detected by MTT,apoptosis test,migration test and invasion test.The levels ofmammalian target of rapamycin(mTOR),phosphorylated mTOR(p-mTOR),eukaryotic initiation factor-4 E(eIF4 E),phosphorylated eIF4 E(p-eIF4 E),ribosomal protein S6 kinase(p70 S6 K),phosphorylated p70 S6 K(p-p70 S6 K),ribosomal protein S6(S6),phosphorylated S6(p-S6),eIF4 E-binding protein(4 E-BP),phosphorylated 4 E-BP(p-4 E-BP)were detected by Western blotting.Results The miR-145 expression levels and cell apoptosis levels between the control group and control+antagonists group showed no significant difference(P>0.05),however,which significantly differed with the miR-145 group and miR-145+antagonists group,and the miR-145 group was the highest,the miR-145+antagonists group was the lowest,and the difference was statistically significant(P<0.05).The cell proliferation,cell migration,cell invasion activities and the expression levels of p-mTOR/mTOR,p-eIF4 E/eIF4 E,p-p70 S6 K/p70 S6 K,p-S6/S6,p-4 E-BP/4 E-BP between the control group and control+antagonists group showed no significant difference(P>0.05),however,which significantly differed with the miR-145 group and miR-145+antagonists group,and the miR-145 group was the lowest,the miR-145+antagonists group was the highest,and the difference was statistically significant(P<0.05).Conclusion miR-145 can regulate the biological characteristics of NSCLC A549 cells through mTOR pathway,promote cell apoptosis,inhibit cell proliferation,migration and invasion.

关 键 词：MIR-145 mTOR途径 非小细胞肺癌 A549细胞 生物学特性 

分 类 号：R734.2[医药卫生—肿瘤]