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微小RNA-125b通过PI3K/Akt/GSK3β信号通路调节人非小细胞肺癌的细胞凋亡。

miRNA-125b regulates apoptosis of human non-small cell lung cancer via the PI3K/Akt/GSK3β signaling pathway.

作者信息

Wang Yingyi, Zhao Ming, Liu Jieying, Sun Zhao, Ni Jianjiao, Liu Hongsheng

机构信息

Oncology Department of Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Dongcheng, Beijing 100730, P.R. China.

Thoracic Surgery Department of China PLA General Hospital, Haidian, Beijing 100853, P.R. China.

出版信息

Oncol Rep. 2017 Sep;38(3):1715-1723. doi: 10.3892/or.2017.5808. Epub 2017 Jul 12.

Abstract

The present investigation demonstrated that regulation of microRNA (miR)-125b affected the apoptosis of human non-small cell lung cancer (NSCLC) through targeting of the PI3K/Akt and Wnt/β-catenin signaling pathways. The expression of miR-125b was assessed in patients with NSCLC, which demonstrated that miR-125b expression in NSCLC tissue was higher than that in para-carcinoma tissue. Furthermore, survival analysis of patients with NSCLC over 3 years indicated that the overall survival (OS) and disease-free survival (DFS) rates of patients with low miR-125b expression were higher than those of patients with high miR-125b expression. Proliferation and apoptosis assays were subsequently conducted in the human NSCLC cell line A549 using MTT assay and Annexin V-FITC/PI kits, respectively. Caspase-3 activity ELISA and western blot analysis were also used to assess caspase-3 activity and the protein expression of Bax, Akt, phosphorylated (p)-Akt, p-GSK3β, Wnt and β-catenin. It was observed that downregulation of miR-125b inhibited the proliferation and induced the apoptosis of A549 cells. Downregulation of miR-125b also suppressed the protein expression of p-Akt, Wnt and β-catenin, and increased caspase-3 activity and Bax protein expression in A549 cells. In addition, downregulation of miR-125b combined with the PI3K inhibitor LY294002 enhanced cell growth inhibition, suppression of p-GSK3β, Wnt and β-catenin protein expression and promotion of caspase-3 activity in A549 cells. These results revealed that the downregulation of miR-125b regulates apoptosis in human NSCLC through the suppression of the PI3K/Akt/GSK3β and Wnt/β-catenin signaling pathways.

摘要

本研究表明,微小RNA(miR)-125b的调控通过靶向PI3K/Akt和Wnt/β-连环蛋白信号通路影响人非小细胞肺癌(NSCLC)的细胞凋亡。对NSCLC患者的miR-125b表达进行评估,结果显示NSCLC组织中miR-125b的表达高于癌旁组织。此外,对NSCLC患者进行的3年以上生存分析表明,miR-125b低表达患者的总生存期(OS)和无病生存期(DFS)率高于miR-125b高表达患者。随后分别使用MTT法和Annexin V-FITC/PI试剂盒在人NSCLC细胞系A549中进行增殖和凋亡检测。还使用Caspase-3活性ELISA和蛋白质印迹分析来评估Caspase-3活性以及Bax、Akt、磷酸化(p)-Akt、p-GSK3β、Wnt和β-连环蛋白的蛋白质表达。结果观察到,miR-125b的下调抑制了A549细胞的增殖并诱导其凋亡。miR-125b的下调还抑制了A549细胞中p-Akt、Wnt和β-连环蛋白的蛋白质表达,并增加了Caspase-3活性和Bax蛋白质表达。此外,miR-125b的下调与PI3K抑制剂LY294002联合使用可增强对A549细胞生长的抑制作用、抑制p-GSK3β、Wnt和β-连环蛋白的蛋白质表达并促进Caspase-3活性。这些结果表明,miR-125b的下调通过抑制PI3K/Akt/GSK3β和Wnt/β-连环蛋白信号通路来调节人NSCLC中的细胞凋亡。

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