MiR-92a targets PTEN/PI3K/Akt pathway to inhibit autophagy in non-small cell lung cancer cells

Aini HU; Yuping WANG; Yibin LU; Mei YANG; Lili CUI; Jian WANG

doi:10.16016/j.1000-5404.201912194

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MiR-92a targets PTEN/PI3K/Akt pathway to inhibit autophagy in non-small cell lung cancer cells

Basic Medicine | 更新时间：2026-01-09

- MiR-92a targets PTEN/PI3K/Akt pathway to inhibit autophagy in non-small cell lung cancer cells
- Journal of Third Military Medical University Vol. 42, Issue 13, Pages: 1301-1307(2020)
- 作者机构：
  
  1.550004 贵阳，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
  2.550004 贵阳，贵州中医药大学第二附属医院检验科
  3.550004 贵阳，贵州省人民医院病理科
- 作者简介：
- 基金信息：
- DOI：10.16016/j.1000-5404.201912194
  CLC： R394.3;R73-36;R734.2
- Received：20 December 2019，
  
  Revised：2020-03-26，
  
  Published：15 July 2020
- 稿件说明：
移动端阅览
Aini HU, Yuping WANG, Yibin LU, et al. MiR-92a targets PTEN/PI3K/Akt pathway to inhibit autophagy in non-small cell lung cancer cells[J]. Journal of Third Military Medical University, 2020, 42(13): 1301-1307.
DOI：

Aini HU, Yuping WANG, Yibin LU, et al. MiR-92a targets PTEN/PI3K/Akt pathway to inhibit autophagy in non-small cell lung cancer cells[J]. Journal of Third Military Medical University, 2020, 42(13): 1301-1307. DOI： 10.16016/j.1000-5404.201912194.

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(

0.001)

p-PI3K、p-Akt阳性细胞率也显著升高(

0.001)

PTEN、LC3-Ⅱ阳性细胞率显著降低(

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(

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(

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(

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(

0.05)

LC3-Ⅱ最弱(

0.05)

miR-92a-inhibitor组p-Akt、p-PI3K蛋白表达最弱(

0.05)

LC3-Ⅱ最强(

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

Objective

To investigate the effect of miR-92a on autophagy in non-small cell lung cancer cells (NSCLC) and its possible mechanism.

Methods

The 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.

Results

The mRNA expression of miR-92a was significantly higher in the NSCLC tissues than the normal lung tissues (

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 (

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 (

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 (

0.01). While opposite phenomena were observe when concerning of the expression of PTEN and LC3-Ⅱ (

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

0.05). And opposite results were seen in the miR-92a-inhibitor group (all

0.05).

Conclusion

MiR-92a targets and inhibits PTEN expression

activates PI3K/Akt signaling pathway

and thus suppresses cell autophagy in NSCLC cells.

关键词

Keywords

references

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