Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

MiR-1224-5p reverses gefitinib resistance in non-small-cell lung cancer cells by modulating RFX5/YAP1/HIF1α axis

Hanxu Tang¹, Chunhua Liu¹, Xiangchun Yu¹, Weiwei Zhao¹, Zexin Gu³, Ying Liu¹, Xin Zheng⁴, Xiangru Meng¹

¹Respiratory Department I, The Second Affiliated Hospital of Qiqihar Medical University, Qiqihar 161000, China; ²Blood Specialty, The Second Affiliated Hospital of Qiqihar Medical University, Qiqihar 161000, China; ³Respiratory Department II, The Second Affiliated Hospital of Qiqihar Medical University, Qiqihar 161000, China; ⁴Digestive Department, The Second Affiliated Hospital of Qiqihar Medical University, Qiqihar 161000, China.

For correspondence:- Xiangru Meng Email: mengxiangru@qmu.edu.cn

Accepted: 2 February 2024 Published: 29 February 2024

Citation: Tang H, Liu C, Yu X, Zhao W, Gu Z, Liu Y, et al. MiR-1224-5p reverses gefitinib resistance in non-small-cell lung cancer cells by modulating RFX5/YAP1/HIF1α axis. Trop J Pharm Res 2024; 23(2):263-272 doi: 10.4314/tjpr.v23i2.5

© 2024 The authors.
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Abstract

Purpose: To investigate the molecular pathways by which miR-1224-5p modulate RFX5/YAP1/HIF1α pathway, thereby promoting gefitinib tolerance within non-small-cell lung cancer (NSCLC).

Methods: To screen differentially expressed miR-1224-5p in NSCLC samples and predict its downstream target gene – RFX5 – by bioinformatics analysis, 60 NSCLC tissues and their corresponding paraneoplastic tissues were collected. Dual-luciferase assays were performed to verify the targeting relationship between miR-1224-5p and RFX5. Four NSCLC cell lines (A549, H1299, H2170, and H1975) and BEAS-2B normal lung epithelial cell lines were used for in vitro experiments. Co-immunoprecipitation (Co-IP) and Western blotting after cycloheximide (CHX) treatment were used to determine the regulatory interaction between YAP1 and HIF1α, with YAP1 modulating HIF1α protein stability.

Results: In NSCLC, downregulation of miR-1224-5p was observed, which resulted in the decrease of RFX5 levels. This reduction in miR-1224-5p levels leads to a decrease in RFX5 levels. Furthermore, restoring miR-1224-5p expression in NSCLC cells made them more sensitive to gefitinib. In vitro, RFX5 elevates YAP1, which in turn boosts the stability of HIF1α. However, miR-1224-5p disrupts this mechanism by influencing the RFX5/YAP1/HIF1α pathway, thus mitigating resistance to gefitinib.

Conclusion: The findings show that miR-1224-5p targets RFX5 to suppress YAP1 transcription, thereby diminishing HIF1α stability and overcoming gefitinib tolerance in NSCLC. These findings identify miR-1224-5p as a promising approach to address gefitinib tolerance in NSCLC.

Keywords: Non-Small Cell Lung Cancer (NSCLC), Gefitinib, miR-1224-5p, RFX5