Effect and mechanism of miR‑146a on malignant biological behaviors of lung adenocarcinoma cell line

Yuan,Fang; Zhang,Suyun; Xie,Wenying; Yang,Sheng; Lin,Tingyan; Chen,Xiangqi

doi:10.3892/ol.2020.11474

Effect and mechanism of miR‑146a on malignant biological behaviors of lung adenocarcinoma cell line

Authors:
- Fang Yuan
- Suyun Zhang
- Wenying Xie
- Sheng Yang
- Tingyan Lin
- Xiangqi Chen
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Affiliations: Department of Respiratory Medicine, Fujian Medical University Union Hospital, Fuzhou, Fujian 350001, P.R. China, Department of Oncology, Fujian Medical University Union Hospital, Fuzhou, Fujian 350001, P.R. China
Published online on: March 23, 2020 https://doi.org/10.3892/ol.2020.11474
Pages: 3643-3652
Copyright: © Yuan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to assess the expression of microRNA‑146a (miR‑146a) in human lung adenocarcinoma cells, its effect on cellular behaviors, and the underlying molecular mechanisms. Reverse transcription‑quantitative PCR (RT‑qPCR) was used to measure miR‑146a expression in the human normal lung epithelial cell line, BEAS‑2B, and human lung adenocarcinoma cell lines, A549, PC‑9 and H1299, to determine whether miR‑146a acts as an oncogene or anti‑oncogene. miR‑146a mimics were transfected into target cells to observe the proliferation, apoptosis, invasion and migration of human lung adenocarcinoma cells. The target genes of miR‑146a were predicted using bioinformatics analysis, and binding sites were validated by dual‑luciferase reporter assay. Target gene expression at the mRNA and protein levels was measured by RT‑qPCR and western blot analysis, respectively. The expression levels of miR‑146a in human lung adenocarcinoma cell lines were lower than its expression in BEAS‑2B (P<0.01). A549 cell line is a EGFR wild‑type lung adenocarcinoma cell line, which is also the most widely studied in NSCLC, and therefore this was chosen as the target cell line for further investigation. Overexpression of miR‑146a in A549 cells can inhibit cell proliferation (P<0.05), promote apoptosis (P<0.05), and reduce the cells' migratory ability (P<0.01). Bioinformatics prediction indicated that interleukin‑1 receptor‑associated kinase 1 (IRAK1) and TNF receptor associated factor 6 (TRAF6) are the target genes of miR‑146a. Dual‑luciferase reporter assay showed that miR‑146a could specifically bind to 3'‑untranslated regions of IRAK1 and TRAF6. The protein and mRNA levels of IRAK1 and TRAF6 were significantly downregulated after miR‑146a overexpression in A549 cells (P<0.01). The results of this study demonstrated that the expression of miR‑146a in human lung adenocarcinoma cells was significantly lower than in normal lung epithelial cells, indicating that miR‑146a acts as an anti‑oncogene. miR‑146a suppresses the proliferation and migration of human lung adenocarcinoma cells by downregulating the expression of IRAK1 and TRAF6.

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