MiR-30e-5p overexpression promotes proliferation and migration of colorectal cancer cells by activating the CXCL12 axis via downregulating PTEN.
10.12122/j.issn.1673-4254.2023.07.04
- Author:
Ke WEI
1
;
Jiwen SHI
1
;
Yuhan XIAO
1
;
Wenrui WANG
2
;
Qingling YANG
3
;
Changjie CHEN
3
Author Information
1. Anhui Provincial Key Laboratory of Cancer Translational Medicine, Bengbu Medical College, Bengbu 233000, China.
2. Department of Biotechnology, Bengbu Medical College, Bengbu 233000, China.
3. Department of Biochemistry and Molecular Biology, Bengbu Medical College, Bengbu 233000, China.
- Publication Type:Journal Article
- Keywords:
CXCL12;
PTEN;
colorectal cancer;
miR-30e-5p
- MeSH:
Humans;
Animals;
Mice;
MicroRNAs/metabolism*;
Cell Line, Tumor;
Cell Proliferation/physiology*;
Mice, Nude;
Cell Movement/physiology*;
Colorectal Neoplasms/pathology*;
Luciferases/metabolism*;
Gene Expression Regulation, Neoplastic;
PTEN Phosphohydrolase/metabolism*;
Chemokine CXCL12/metabolism*
- From:
Journal of Southern Medical University
2023;43(7):1081-1092
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the regulatory effects of miR-30e-5p on biological behaviors of colorectal cancer cells and the role of PTEN/CXCL12 axis in mediating these effects.
METHODS:Bioinformatic analysis was performed to explore the differential expression of miR-30e-5p between colorectal cancer tissues and normal tissues. RT-qPCR was used to detect the differential expression of miR-30e-5p in intestinal epithelial cells and colorectal cancer cells. Bioinformatics and dual luciferase assay were used to predict and validate the targeting relationship between miR-30e-5p and PTEN. Human and murine colorectal cancer cell lines were transfected with miR-30e-5p mimics, miR-30e-5p inhibitor, miR-30e-5p mimics+LV-PTEN, or miR-30e-5p inhibitor + si-PTEN. The changes in biological behaviors of the cells were detected using plate clone formation assay, CCK-8 assay, flow cytometry, scratch healing and Transwell assays. PTEN and CXCL12 expressions in the cancer cells were detected by Western blotting. The effects of miR-30e-5p inhibitor on colorectal carcinogenesis and development were observed in nude mice.
RESULTS:Bioinformatic analysis showed that miR-30e-5p expression was significantly elevated in colorectal cancer tissues compared with the adjacent tissue (P < 0.01). Higher miR-30e-5p expression was detected in colorectal cancer cell lines than in intestinal epithelial cells (P < 0.01). Dual luciferase assay confirmed the targeting relationship between miR-30e-5p and PTEN (P < 0.05). Transfection with miR-30e-5p mimics significantly enhanced proliferation and metastasis and inhibited apoptosis of the colorectal cancer cells (P < 0.05), and co-transfection with LV-PTEN obviously reversed these changes (P < 0.05). MiR-30e-5p mimics significantly inhibited PTEN expression and enhanced CXCL12 expression in the cancer cells (P < 0.01), and miR-30e-5p inhibitor produced the opposite effect. Transfection with miR-30e-5p inhibitor caused cell cycle arrest in the cancer cells, which was reversed by co-transfection with si-PTEN (P < 0.05). In the in vivo experiments, the colorectal cancer cells transfected with miR-30e-5p inhibitor showed significantly lowered tumorigenesis.
CONCLUSION:Overexpression of miR-30e-5p promotes the malignant behaviors of colorectal cancer cells by downregulating PTEN to activate the CXCL12 axis.
