Effect of miR-325-3p targeting PRELID1 gene in regulation of EMT pathway on invasion and migration of colon cancer cells and their mechanisms
10.13481/j.1671-587X.20250504
- VernacularTitle:miR-325-3p靶向PRELID1基因介导EMT通路对结肠癌细胞侵袭和迁移的影响及其机制
- Author:
Pingsheng ZHU
1
;
Sitang GE
;
Lugen ZUO
;
Deli CHEN
;
Yangyang ZHANG
Author Information
1. 蚌埠医科大学第一附属医院胃肠外科,安徽 蚌埠 233000
- Keywords:
MicroRNA-325-3p;
Relevant evolutionary and lymphoid interest domain containing protein 1;
Colon neoplasm;
Cell invasion;
Cell migration;
Epithelial-mesenchymal transition
- From:
Journal of Jilin University(Medicine Edition)
2025;51(5):1185-1193
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of microRNA-325-3p(miR-325-3p)over-expression on the invasion and migration of colon cancer cells,and to clarify their mechanisms.Methods:The tumor tissue and the corresponding adjacent normal tissue of 25 patients clearly diagnosed with colon cancer were collected.The expression levels of miR-325-3p and relevant evolutionary and lymphoid interest domain containing protein 1(PRELID1)mRNA were detected by real-time fluorescence quantitative PCR(RT-qPCR)method.The expression levels of miR-325-3p and PRELID1 mRNA and the expression levels of PRELID1 protein in human normal colon cells NCM460 and colon cancer cells SW480,HCT116 and HT-29 were detected by RT-qPCR and Western blotting methods.The SW480 cells were divided into control group,NC-mimics group,miR-325-3p mimics group,miR-325-3p mimics+oe-NC group and miR-325-3p mimics+oe-PRELID1 group.The invasion and migration abilities of cells in various groups were detected by Transwell chamber assay and scratch healing assay,respectively.The expression levels of E-Cadherin,N-Cadherin and Vimentin in the epithelial-mesenchymal transition(EMT)of cells in various groups were detected by Western blotting method.The downstream target genes of miR-325-3p were predicted using the Targetscan bioinformatics website,and the targeted regulatory relationship between miR-325-3p and PRELID1 was verified by the dual-luciferase assay.Results:Compared with adjacent normal tissue,the expression level of miR-325-3p in cancer tissue was significantly decreased(P<0.05),while the expression level of PRELID1 mRNA was significantly increased(P<0.05).The expression levels of miR-325-3p and PRELID1 mRNA was negatively correlated in colon cancer tissue(r<0,R2=0.392,P<0.001).Compared with NCM460 cells,the expression levels of miR-325-3p in SW480,HCT116 and HT-29 cells were significantly decreased(P<0.05),and the expression levels of PRELID1 mRNA were significantly increased(P<0.05).Compared with control group and NC-mimics group,the number of invasive SW480 cells in miR-325-3p mimics group was significantly reduced(P<0.05),the rate of scratch healing was decreased(P<0.05),and the expression level of E-Cadherin protein was increased,while the expression levels of N-Cadherin and Vimentin proteins were decreased(P<0.05).The dual-luciferase assay confirmed that PRELID1 was the direct target of miR-325-3p.Compared with miR-325-3p mimics+oe-NC group,the number of invasive SW480 cells in miR-325-3p mimics+oe-PRELID1 group was significantly increased(P<0.05),the rate of scratch healing was increased(P<0.05),the expression level of E-Cadherin protein was decreased,and the expression levels of N-Cadherin and Vimentin proteins were increased(P<0.05).Conclusion:Over-expression of miR-325-3p can inhibit EMT process by down-regulating PRELID1 expression,thereby inhibiting SW480 cell invasion and migration.