MiR-224-5p regulates chemoresistance in colorectal cancer via Bcl-2-mediated autophagy.
10.11817/j.issn.1672-7347.2025.240243
- Author:
Hui ZHOU
1
;
Meng WU
2
;
Shaihong ZHU
3
;
Yi ZHANG
4
Author Information
1. Department of Gastrointestinal Surgery, Third Xiangya Hospital, Central South University, Changsha 410013, China. oneonedreame@gmail.com.
2. Department of Dermatology, Hunan Provincial People's Hospital, Changsha 410002, China.
3. Department of Gastrointestinal Surgery, Third Xiangya Hospital, Central South University, Changsha 410013, China.
4. Department of Gastrointestinal Surgery, Affiliated Hospital of Xuzhou Medical University, Xuzhou 221002, China. drzhy@aliyun.com.
- Publication Type:Journal Article
- Keywords:
B-cell lymphoma-2;
autophagy;
chemoresistance;
colorectal cancer;
miR-224-5p
- MeSH:
Humans;
MicroRNAs/physiology*;
Colorectal Neoplasms/drug therapy*;
Drug Resistance, Neoplasm/genetics*;
Autophagy/drug effects*;
Fluorouracil/pharmacology*;
Oxaliplatin;
Cell Line, Tumor;
Proto-Oncogene Proteins c-bcl-2/metabolism*;
Gene Expression Regulation, Neoplastic
- From:
Journal of Central South University(Medical Sciences)
2025;50(2):190-203
- CountryChina
- Language:English
-
Abstract:
OBJECTIVES:Oxaliplatin (OXA) and 5-fluorouracil (5-FU) are 2 commonly used chemotherapeutic agents for colorectal cancer (CRC). MicroRNAs (miRNAs, miRs) play crucial roles in the development of chemoresistance in various cancers. However, the role and mechanism of miR-224-5p in regulating CRC chemoresistance remain unclear. This study aims to investigate the function of miR-224-5p in chemoresistant CRC cells and the underlying mechanisms.
METHODS:CRC datasets GSE28702 and GSE69657 were downloaded from the Gene Expression Omnibus (GEO) database. Differentially expressed miRNAs between drug-sensitive and resistant groups (OXA or 5-FU) were analyzed, and miR-224-5p was identified as the target miRNA. Chemoresistant cell lines HCT15-OXR, HCT15-5-FU, SW480-OXR, and SW480-5-FU were established. Transient transfections were performed using miR-224-5p mimics, inhibitors, and their respective negative controls (control mimic, control inhibitor) in these cell lines. Cells were treated with different concentrations of OXA or 5-FU post-transfection, and the half-maximal inhibitory concentration (IC50) was determined using the cell counting kit-8 (CCK-8) assay. Cell proliferation was assessed by CCK-8 and colony formation assays. The expression levels of miR-224-5p, LC3, and P62 were measured by real-time polymerase chain reaction (real-time PCR) and/or Western blotting. Autophagic flux was assessed using a tandem fluorescent-tagged LC3 reporter assay. TargetScan 8.0, miRTarBase, miRPathDB, and HADb were used to predict B-cell lymphoma-2 (Bcl-2) as a potential miR-244-5p target, which was further validated by dual-luciferase reporter assays.
RESULTS:Chemoresistant CRC cells exhibited down-regulated miR-224-5p expression, whereas up-regulation of miR-224-5p enhanced chemotherapy sensitivity. Exposure to OXA or 5-FU significantly increased autophagic activity in chemoresistant CRC cells, which was reversed by miR-224-5p overexpression. Dual-luciferase assays verified Bcl-2 as a direct target of miR-224-5p.
CONCLUSIONS:MiR-224-5p regulates chemoresistance in CRC by modulating autophagy through direct targeting of Bcl-2.
