miR-15a 在局部低氧微环境中非小细胞肺癌 A549 细胞中的表达分析。

MiR-15a expression analysis in non-small cell lung cancer A549 cells under local hypoxia microenvironment.

机构信息

Department of Respiratory Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, P.R. China.

出版信息

Eur Rev Med Pharmacol Sci. 2017 May;21(9):2069-2074.

PMID:28537679

Abstract

OBJECTIVE

Lung cancer is a common tumor in the clinic. Hypoxia is an important biological characteristic in the solid malignant tumor. MiRNA participates in cell proliferation, differentiation, and apoptosis. This study tested hypoxia-inducible factor 1-α (HIF-1α) in lung cancer patients and analyzed the microRNA-15a (miR-15a) expression in A549 cells under different local hypoxia microenvironments.

PATIENTS AND METHODS

A total of 40 non-small cell lung cancer (NSCLC) patients in First Affiliated Hospital of Zhengzhou University between Jan 2015 and Jan 2016 were involved in this study. The serum and tissue samples of lung cancer were collected. Serum HIF-1α level was tested by enzyme-linked immunosorbent assay (ELISA) assay. HIF-1α expression in tissue was evaluated by using the immunohistochemistry. A549 cells were cultured under normoxic, hypoxic, and anaerobic environment, respectively. HIF-1α mRNA and miR-15a levels were determined by RT-PCR.

RESULTS

HIF-1α levels were up-regulated in serum and tissue (p<0.05). HIF-1α mRNA increased, while miR-15a down-regulated in A549 from hypoxia and anaerobic groups compared with control (p<0.05). HIF-1α shRNA transfection significantly reduced HIF-1α and elevated miR-15a level (p<0.05). MiR-15a shRNA transfection exhibited no statistical impact on HIF-1α expression (p>0.05).

CONCLUSIONS

HIF-1α highly expressed in lung cancer patients. MiR-15a levels were down-regulated in A549 cells under hypoxia and anaerobic conditions.

摘要

目的

肺癌是临床上常见的肿瘤。缺氧是实体恶性肿瘤的重要生物学特征。miRNA 参与细胞增殖、分化和凋亡。本研究检测了肺癌患者中缺氧诱导因子 1-α（HIF-1α），并分析了不同局部缺氧微环境下 A549 细胞中 microRNA-15a（miR-15a）的表达。

患者与方法

本研究共纳入 2015 年 1 月至 2016 年 1 月郑州大学第一附属医院收治的 40 例非小细胞肺癌（NSCLC）患者。采集肺癌患者血清和组织标本，采用酶联免疫吸附试验（ELISA）检测血清 HIF-1α 水平，免疫组织化学法检测组织中 HIF-1α 的表达。分别在常氧、缺氧和厌氧环境下培养 A549 细胞，采用 RT-PCR 检测 HIF-1αmRNA 和 miR-15a 水平。

结果

血清和组织中 HIF-1α 水平升高（p<0.05）。与对照组相比，缺氧和厌氧组 A549 细胞 HIF-1αmRNA 增加，miR-15a 降低（p<0.05）。HIF-1αshRNA 转染后 HIF-1α 明显降低，miR-15a 水平升高（p<0.05）。miR-15a shRNA 转染对 HIF-1α 表达无统计学影响（p>0.05）。