Molecular Mechanism of AP000892.6 Affecting Bladder Cancer Cells Migration,Invasion and Epithelial-mesenchymal Transition by Targe-ting MiR-616-5p
10.11969/j.issn.1673-548X.2023.12.007
- VernacularTitle:AP000892.6通过靶向miR-616-5p影响膀胱癌细胞迁移、侵袭和上皮间充质转化的分子机制
- Author:
Jinhui YANG
1
;
Jiantao SUN
;
Xiaohui LI
Author Information
1. 471000 郑州大学附属洛阳中心医院泌尿外科
- Keywords:
Bladder neoplasms;
AP000892.6;
MiR-616-5p;
Epithelial-mesenchymal transition
- From:
Journal of Medical Research
2023;52(12):26-31
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of long-chain non-coding RNA(lncRNA)AP000892.6 on the migration,inva-sion and epithelial-mesenchymal transition of bladder cancer cells and the related molecular mechanism.Methods The expression of AP000892.6 in bladder cancer and adjacent tissues was analyzed using GEPIA dataset.The expression levels of AP000892.6 in bladder cancer cell lines 647V,T24,UMUC3,5637,RT4 and normal bladder epithelial cells SV-HUC-1 were detected by real-time quanti-tative polymerase chain reaction(RT-qPCR).The AP000892.6 overexpression plasmid or negative control plasmid was transfected into T24 cells,and the experiment was divided into AP000892.6 group and NC group.The plasmid transfection efficiency was verified by RT-qPCR.The migration and invasion abilities of transfected T24 cells were detected by cell scratch method and Transwell assay,re-spectively.The downstream target gene of AP000892.6 was predicted and validated using bioinformatics method and dual-luciferase re-porter gene experiment.The expression of AP000892.6downstream target gene was detected by RT-qPCR.Western blot was used to de-tect the expression of epithelial phenotype proteins E-cadherin,ZO-1 and mesenchymal phenotype proteins Vimentin,N-cadherin and Twist.Results Compared with adjacent tissues,the expression of AP000892.6 was decreased in bladder cancer tissues(P＜0.01).Compared with SV-HUC-1 cells,the expression of AP000892.6 was decreased in bladder cancer cell lines(P＜0.05),and the rela-tive expression level of AP000892.6 in T24 cells was the lowest(P＜0.01).Compared with the NC group,overexpression of AP000892.6 inhibited the migration(P＜0.01)and invasive ability(P＜0.01)of bladder cancer T24 cells.Dual-luciferase reporter gene analysis proved that miR-616-5p was the target gene of AP000892.6(P＜0.01).Compared with the NC group,AP000892.6 negatively regu-lated the expression of miR-616-5p(P＜0.01).Compared with the NC group,the expression of epithelial phenotype proteins E-cad-herin and ZO-1 were up-regulated in T24 cells of AP000892.6group,and the expression of mesenchymal phenotype proteins Vimentin,N-cadherin and Twist were down-regulated.Conclusion AP000892.6 inhibits the migration,invasion and epithelial-mesenchymal transition process of bladder cancer T24 cells by targeting miR-616-5p.
