|Table of Contents|

miRNA-139-5p reverses cisplatin resistance in non-small cell lung cancer cells A549 through targeted regulation of CXCR4/CXCL12 signaling pathway

Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:
2023 08
Page:
1379-1385
Research Field:
Publishing date:

Info

Title:
miRNA-139-5p reverses cisplatin resistance in non-small cell lung cancer cells A549 through targeted regulation of CXCR4/CXCL12 signaling pathway
Author(s):
CHEN Jialiang1; ZHOU Xiangdong1; 2; 3; LIU Chang1; LI Qi1; HUANG Huaping1; HAN Zhong1; CHEN Xiaomei4
1.Department of Respiratory Medicine;4.Department of General Medicine,the First Affiliated Hospital of Hainan Medical College,Hainan Haikou 571199,China;2.Key Laboratory of Emergency and Trauma,Ministry of Education,Hainan Haikou 571199,China;3.Innovative Unit of Island Emergency Medicine,Chinese Academy of Medical Sciences,Hainan Haikou 571199,China.
Keywords:
miR-139-5p; non-small cell lung cancer; CXCR4/CXCL12 signaling pathway; cisplatin resistance
PACS:
R734.2
DOI:
10.3969/j.issn.1672-4992.2023.08.001
Abstract:
Objective:To explore the effect of miRNA-139-5p on cisplatin (DDP) resistance in non-small cell lung cancer cells A549 and its possible molecular mechanism.Methods:DDP concentration increasing method was used to induce A549 cells to establish DDP-resistant cell line A549/DDP.The miR-139-5p mimics,nonsense sequence (mimics-NC),CXCR4 overexpression plasmid (pcDNA3.1-CXCR4),and pcDNA3.1 empty plasmid (Vector) were transfected into A549/DDP cells.Then the cells were divided into mimics-NC group (transfected mimics),mimics+Vector group (cotransfected with miR-139-5p mimics and Vector plasmid),and mimics+CXCR4 group（cotransfected with miR-139-5p mimics and CXCR4 overexpression plasmid）.A blank control group (blank) was set.After treatment with different concentrations of DDP,MTT method was used to detect the cell proliferation activity,and IC50 value and drug resistance index (RI) were calculated.qRT-PCR method was used to detect the expression levels of miR-139-5p and CXCR4 mRNA in cells.Flow cytometry was used to detect cell apoptosis rate.Western blot method was used to detect the expression of drug resistance-related protein P-glycoprotein (P-gp),multidrug resistance-related protein 1 (MRP1) and CXCR4/CXCL12 signaling pathway related proteins in cells.Dual-luciferase reporter gene experiment verified the targeting relationship between miR-139-5p and CXCR4.Results:After 24 hours of treatment with different concentrations of DDP,the IC50 values of the drug-resistant strain A549/DDP and its parent A549 cells were (208.87±27.89) μmol/L and (31.66±6.30) μmol/L,with RI=6.59.Compared with the parental A549 cells,the expression level of miR-139-5p in resistant strain A549/DDP cells was significantly reduced (P＜0.05),while the expression levels of CXCR4 mRNA and protein were significantly increased (P＜0.05).Compared with the the mimics-NC group,the proliferation activity of A549/DDP cells in the mimics group was significantly reduced (P＜0.05),and the apoptosis rate was significantly increased (P＜0.05),and the expression level of CXCR4 mRNA and protein in the cells,as well as P-gp,MRP1,PI3K (p110α) and p-AKT/AKT were markedly reduced (P＜0.05).Compared with the mimics+Vector group,the proliferation activity of A549/DDP cells in the mimics+CXCR4 group was significantly increased (P＜0.05),and the apoptosis rate was markedly reduced (P＜0.05),while the expression levels of CXCR4,P-gp,MRP1,PI3K (p110α) and p-AKT proteins were significantly increased (P＜0.05).The dual-luciferase reporter gene experiment confirmed that miR-139-5p targeted and negatively regulated CXCR4 expression.Conclusion:miRNA-139-5p improves the drug sensitivity of non-small cell lung cancer DDP resistant cell lines A549/DDP to DDP by targeting down-regulation of CXCR4 expression.

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Memo

Memo:
National Natural Science Foundation of China(No.82011530049）；国家自然科学基金项目（编号：82011530049）；海南省卫生计生行业科研项目（编号：18A200141）
Last Update: 1900-01-01