[1]胡爱妮,王豫萍,路毅斌,等.miR-92a靶向PTEN/PI3K/Akt通路抑制非小细胞肺癌细胞的自噬[J].陆军军医大学学报(原第三军医大学学报),2020,42(13):1301-1307.
 HU Aini,WANG Yuping,LU Yibin,et al.MiR-92a targets PTEN/PI3K/Akt pathway to inhibit autophagy in non-small cell lung cancer cells[J].J Amry Med Univ (J Third Mil Med Univ),2020,42(13):1301-1307.
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miR-92a靶向PTEN/PI3K/Akt通路抑制非小细胞肺癌细胞的自噬(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
42卷
期数:
2020年第13期
页码:
1301-1307
栏目:
基础医学
出版日期:
2020-07-15

文章信息/Info

Title:
MiR-92a targets PTEN/PI3K/Akt pathway to inhibit autophagy in non-small cell lung cancer cells
作者:
胡爱妮王豫萍路毅斌杨梅崔丽丽王剑
 
贵州医科大学医学检验学院临床微生物学及免疫学教研室;贵州中医药大学第二附属医院检验科;贵州省人民医院病理科
Author(s):
HU Aini WANG Yuping LU Yibin YANG Mei CUI Lili WANG Jian

Department of Clinical Microbiology and Immunology, School of Laboratory Medicine, Guizhou Medical University, Guiyang, Guizhou Province, 550004; 2Department of Clinical Laboratory, the Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou Province, 550004; 3Department of Pathology, Guizhou People’s Hospital, Guiyang, Guizhou Province, 550004

关键词:
非小细胞肺癌miR-92aPTENPI3K/Akt信号通路自噬
Keywords:
 
分类号:
R394.3; R73-36; R734.2
文献标志码:
A
摘要:

目的探讨miR-92a对非小细胞肺癌(non-small cell lung cancer, NSCLC)细胞自噬的影响及其可能的作用机制。方法收集2016年1月至2018年12月贵州中医药大学第二附属医院收治的53例NSCLC患者的癌组织和癌旁肺组织标本,RT-PCR和免疫组化检测癌组织及癌旁组织内miR-92a、PTEN、p-PI3K、p-Akt和LC3-Ⅱ表达。将miR-92a-mimics(类似物)、miR-92a-NC(抑制剂对照)和miR-92a-inhibitor(抑制物)分别转染至A549细胞,48 h后吖啶橙染色检测细胞自噬,双荧光素酶报告实验验证miR-92a与PTEN的靶向关系,Western blot检测各转染组细胞内PTEN、p-PI3K、p-Akt和LC3-Ⅱ的蛋白表达。结果与癌旁肺组织比较,肺癌组织中miR-92a mRNA的表达显著升高(P<0.001),p-PI3K、p-Akt阳性细胞率也显著升高(P<0.001),PTEN、LC3-Ⅱ阳性细胞率显著降低(P<0.001)。miR-92a、PTEN和LC3-Ⅱ在不同性别、年龄中的表达差异无统计学意义(P>0.05),在存在淋巴结转移、TNM Ⅲ~Ⅳ期及分化程度较低的病灶中,miR-92a高表达者显著多于无淋巴结转移、TNMⅠ~Ⅱ期及分化程度较高的病灶(P<0.01);在存在淋巴结转移、TNMⅢ~Ⅳ期及分化程度较低的病灶中,PTEN和LC3-Ⅱ高表达者显著低于无淋巴结转移、TNMⅠ~Ⅱ期及分化程度较高的病灶(P<0.01)。miR-92a-mimics组细胞啶橙染色荧光最弱,miR-92a-inhibitor组最强。3组中miR-92a-mimics组p-Akt、p-PI3K蛋白表达最高(P<0.05),LC3-Ⅱ最弱(P<0.05),miR-92a-inhibitor组p-Akt、p-PI3K蛋白表达最弱(P<0.05),LC3-Ⅱ最强(P<0.05)。结论miR-92a在非小细胞肺癌中通过靶向抑制PTEN表达,刺激PI3K/Akt信号通路活性,从而抑制细胞的自噬。

Abstract:

ObjectiveTo investigate the effect of miR-92a on autophagy in non-small cell lung cancer cells (NSCLC) and its possible mechanism. MethodsThe paired NSCLC tissues and paracancerous tissues were collected from 53 NSCLC patients treated in the Second Affiliated Hospital of Guizhou University of Traditional Chinese Medicine from January 2016 to December 2018. The expression levels of miR-92a and PTEN, p-PI3K, p-Akt and LC3-Ⅱ in the paired tissues were detected by RT-PCR and immunohistochemical assay. MiR-92a-mimics, miR-92a-NC and miR-92a-inhibitor were respectively transfected into A549 cells. After 48 h, acridine orange staining was used to detect autophagy. Dual luciferase reporter assay was employed to verify the targeting relationship between miR-92a and PTEN, and Western blotting was performed to detect the expression of PTEN, p-PI3K, p-Akt and LC3-Ⅱ in each transfected cells. ResultsThe mRNA expression of miR-92a was significantly higher in the NSCLC tissues than the normal lung tissues (P<0.001). The percentages of p-PI3K and p-Akt positive cells were obviously higher, while those of LC3-Ⅱ and PTEN positive cells were notably lower in the cancer tissues than the normal lung tissues (P<0.001). There were no statistical differences in the expression levels of miR-92a, PTEN and LC3-Ⅱ among the patients of different genders and ages (P>0.05). However, for the patients with lymph node metastasis, at TNM Ⅲ~Ⅳ stage and lower differentiation, the expression levels of miR-92a in the cancer tissue was significantly higher when compared with those without metastasis, at TNM Ⅰ~Ⅱ stage and higher differentiation (P<0.01). While opposite phenomena were observe when concerning of the expression of PTEN and LC3-Ⅱ (P<0.01). Pyridine orange staining showed that the intracellular fluorescence was the weakest in the A549 cells transfected with miR-92a-mimics, while strongest in those with miR-92a-inhibitor. Among the 3 groups of cells with transfection, the expression of p-Akt and p-PI3K protein was the highest, and that of LC3-Ⅱ was the lowest in the miR-92a-mimics group (all P<0.05). And opposite results were seen in the miR-92a-inhibitor group (all P<0.05). ConclusionMiR-92a targets and inhibits PTEN expression, activates PI3K/Akt signaling pathway, and thus suppresses cell autophagy in NSCLC cells.

更新日期/Last Update: 2020-07-06