Overexpression of microRNA‑124‑5p sensitizes non‑small cell lung cancer cells to treatment with 5‑fluorouracil via AEG‑1 regulation

Tan,Xiaoxia; Zhang,Chuancui; Gao,Weidong; Sun,Bei; Jiang,Baozhen; Song,Peng

doi:10.3892/ol.2020.12266

Overexpression of microRNA‑124‑5p sensitizes non‑small cell lung cancer cells to treatment with 5‑fluorouracil via AEG‑1 regulation

Authors:
- Xiaoxia Tan
- Chuancui Zhang
- Weidong Gao
- Bei Sun
- Baozhen Jiang
- Peng Song
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Affiliations: Department of Respiration, The Third People's Hospital of Linyi City, Linyi, Shandong 371312, P.R. China
Published online on: November 3, 2020 https://doi.org/10.3892/ol.2020.12266
Article Number: 5

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Abstract

Chemotherapeutic resistance represents a major obstacle for the treatment of patients with non‑small cell lung cancer (NSCLC); however, the associated molecular mechanisms underpinning the development of resistance remain poorly characterized. In the current study, 5‑fluorouracil (5‑FU)‑resistant A549 cells (A549/5‑FU) were generated from A549 cells. Reverse transcription‑quantitative PCR and western blotting were used to detect microRNA(miR)‑124‑5p and astrocyte elevated gene 1 (AEG‑1) expression levels in cells and tumor tissues. In addition, the cytotoxic effect of 5‑FU on the cells was determined using the Cell Counting Kit‑8 assay, and the Dual‑luciferase reporter assay was used to validate AEG‑1 as a target gene of miR‑124‑5p. Transfection with a miR‑124‑5p mimic enhanced inhibition of cell viability induced by 5‑FU in A549/5‑FU cells, whereas miR‑124‑5p inhibitor transfection partially reversed 5‑FU‑induced cell viability inhibition in A549 and H1299 cells. A decrease in miR‑124‑5p expression level was observed in A549/5‑FU cells compared with the parental A549 cells. Furthermore, AEG‑1 was predicted as a target gene of miR‑124‑5p, and its expression was increased in A549/5‑FU cells compared with A549 cells. Additionally, the upregulation of miR‑124‑5p was associated with lower expression levels of AEG‑1 in A549/5‑FU cells, compared with parental A549 cells. Moreover, the Dual‑luciferase reporter assay confirmed the ability of miR‑124‑5p to bind directly to the 3'‑untranslated region of AEG‑1 mRNA. Notably, the overexpression of AEG‑1 reversed the ability of the miR‑124‑5p mimic to increase the sensitivity of A549/5‑FU cells to 5‑FU treatment. Additionally, a significant negative correlation between miR‑124‑5p expression and AEG‑1 mRNA levels was detected in 40 pairs of NSCLC tissues and their corresponding adjacent paracancerous tissues. The results of the present study indicated that miR‑124‑5p may regulate the chemotherapeutic sensitivity of NSCLC cells, and may therefore represent a promising biomarker or therapeutic target for patients with NSCLC.

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