miR-205作为非小细胞肺癌的生物标志物

miR-205 as a biological marker in non-small cell lung cancer.

作者信息

Duan Baoqi, Guo Tao, Sun Haoliang, Cai Rui, Rui Qinglin, Xi Zhaoqing

机构信息

Nanjing University of Chinese Medicine, 210029, China; Emergency Department of Jiangsu Province, Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine, 210029, China.

Department of Cardiothoracic Surgery, Jiangsu Province People's Hospital Affiliated to Nanjing Medical University, 210029, China.

出版信息

Biomed Pharmacother. 2017 Jul;91:823-830. doi: 10.1016/j.biopha.2017.04.086. Epub 2017 May 23.

DOI:10.1016/j.biopha.2017.04.086

PMID:28501009

Abstract

OBJECTION

The aim of this study is to explain the significance and mechanism of miR-205 in the diagnosis and treatment of non-small cell lung cancer.

METHODS

The 70 advanced NSCLC patients, treated in our hospital, were collected from 2011.10 to 2013.9, taking the tissues from cancer and adjacent tissues to measure the miR-205 expression, evaluate the AKT gene and protein expression of cancer and adjacent normal tissues by RT-PCR and Immunohistochemistry (IHC) assays and analyzing the correlation between miR-205 and AKT. Following up the patients for 2 years; Recording patients' survival time. In the cell experiment, Selecting A549 cell as research object, the cells were divided into three groups: Normal control group (NC), Blank control group (BL) and si-miR-205 transfection group (si-miR-205). Cell proliferation rate and apoptosis rate were detected by MTT method and flow cytometry; Measuring invasion and migration of difference groups by transwell and scratch testing, measured the Akt, mTOR,P21, MMP2 and MMP9 gene expression and detected Akt, p-Akt, mTOR, p-mTOR, P21, MMP2 and MMP 9 protein expression levels.

RESULTS

Compared with adjacent normal tissue, the miR-205 and AKT gene expression level was significantly increased in NSCLC tissues (P<0.05) and the AKT protein expression was stronger than that of healthy tissues, miR-205 was positive correlation with AKT; In the overall survival, MiR-205 high expression group was significantly higher than low expression group (P<0.05). In the cell experiment, Compared with NC and BL groups, si-miR-205 could significantly reduced the biological activity of A549 cells in proliferation, invasion and migration, and promoted the apoptosis of A549 cells (P<0.05, respectively). Akt, p-Akt, mTOR, p-mTOR, P21, MMP 2 and MMP 9 gene and protein expression of si-miR-205 group were significantly compared with NC and BL groups (P<0.05, respectively).

CONCLUSION

miRNA-205 might serve as a potential biomarker for the prognosis of advanced NSCLC, and inhibiting miR-205 expression could decrease A549 cells biological activity by regulating Akt/mTOR/P21 and Akt/MMP 2/MMP 9signaling pathway.

摘要

目的

本研究旨在阐述miR-205在非小细胞肺癌诊断和治疗中的意义及机制。

方法

收集2011年10月至2013年9月在我院接受治疗的70例晚期非小细胞肺癌患者，取癌组织及癌旁组织检测miR-205表达，采用逆转录聚合酶链反应（RT-PCR）和免疫组织化学（IHC）法检测癌组织及癌旁正常组织中AKT基因及蛋白表达，并分析miR-205与AKT的相关性。对患者进行2年随访，记录患者生存时间。细胞实验中，选取A549细胞为研究对象，分为三组：正常对照组（NC）、空白对照组（BL）和si-miR-205转染组（si-miR-205）。采用MTT法和流式细胞术检测细胞增殖率和凋亡率；通过Transwell和划痕实验检测不同组细胞的侵袭和迁移能力，检测Akt、mTOR、P21、MMP2和MMP9基因表达，并检测Akt、p-Akt、mTOR、p-mTOR、P21、MMP2和MMP9蛋白表达水平。

结果

与癌旁正常组织相比，非小细胞肺癌组织中miR-205和AKT基因表达水平显著升高（P<0.05），AKT蛋白表达强于健康组织，miR-205与AKT呈正相关；在总生存方面，miR-205高表达组显著高于低表达组（P<0.05）。细胞实验中，与NC组和BL组相比，si-miR-205可显著降低A549细胞在增殖、侵袭和迁移方面的生物学活性，并促进A549细胞凋亡（P均<0.05）。si-miR-205组Akt、p-Akt、mTOR、p-mTOR、P21、MMP2和MMP9基因及蛋白表达与NC组和BL组相比均显著降低（P均<0.05）。