Journal Article

Enhancing RECK Expression Through miR-21 Inhibition: A Promising Strategy for Bladder Carcinoma Control

Romão, Poliana, Maluf, Feres Camargo, Nahas, William, Guimarães, Vanessa Ribeiro, dos Santos, Gabriel Arantes, Silva, Iran, Candido, Patrícia, Viana, Nayara Izabel, Leite, Katia Ramos Moreira, da Silva Gomes, Paulo Ricardo, Reis, Sabrina T., de Camargo, Juliana Alves, Pimenta, Ruan, dos Santos, Paulo Rodolfo Moraes, Gonçalves, Guilherme Lopes

2025

Overview

Bladder carcinoma (BC) is the tenth most frequent malignancy worldwide, with high morbidity and mortality rates. Despite recent treatment advances, high-grade BC and muscle-invasive BC present with significant progression and recurrence rates, urging the need for alternative treatments. The microRNA-21 (miR-21) has superexpression in many malignancies and is associated with cellular invasion and progression. One of its mechanisms of action is the regulation of RECK, a tumor suppressor gene responsible for inhibiting metalloproteinases, including MMP9. In a high-grade urothelial cancer cell line, we aimed to assess if miR-21 downregulation would promote RECK expression and decrease MMP9 expression. We also evaluated cellular migration and proliferation potential by inhibition of this pathway. In a T24 cell line, we inhibited miR-21 expression by transfection of a specific microRNA inhibitor (anti-miR-21). There were also control and scramble groups, the last with a negative microRNA transfected. After the procedure, we performed a genetic expression analysis of miR-21, RECK, and MMP9 through qPCR. Migration, proliferation, and protein expression were evaluated via wound healing assay, colony formation assay, flow cytometry, and immunofluorescence.After anti-miR-21 transfection, miR-21 expression decreased with RECK upregulation and MMP9 downregulation. The immunofluorescence assay showed a significant increase in RECK protein expression ( p  < 0.0001) and a decrease in MMP9 protein expression ( p  = 0.0101). The anti-miR-21 transfection significantly reduced cellular migration in the wound healing assay ( p  < 0.0001). Furthermore, in the colony formation assay, the anti-miR-21 group demonstrated reduced cellular proliferation ( p  = 0.0008), also revealed in the cell cycle analysis by flow cytometry ( p  = 0.0038). Our results corroborate the hypothesis that miR-21 is associated with BC cellular migration and proliferation, revealing its potential as a new effective treatment for this pathology.

• Publisher: Springer US,Springer Nature B.V
• Subject: Assaying / Biochemistry / Biomedical and Life Sciences / Biomedicine / Bladder / Bladder cancer / Cancer / Carcinoma / Cell cycle / Cell Line, Tumor / cell lines / Cell migration / Cell Movement / Cell Proliferation / Down-regulation / Flow cytometry / fluorescent antibody technique / Gelatinase B / Gene Expression Regulation, Neoplastic / Genetic analysis / GPI-Linked Proteins - biosynthesis / GPI-Linked Proteins - genetics / GPI-Linked Proteins - metabolism / Human Genetics / Humans / Immunofluorescence / Malignancy / Matrix Metalloproteinase 9 - genetics / Matrix Metalloproteinase 9 - metabolism / Medical Microbiology / metalloproteinases / microRNA / MicroRNAs / MicroRNAs - antagonists & inhibitors / MicroRNAs - genetics / MicroRNAs - metabolism / miRNA / Morbidity / mortality / neoplasm cells / Original Article / Protein expression / protein synthesis / Proteins / Ribonucleic acid / RNA / Transfection / Tumor suppressor genes / Urinary Bladder Neoplasms - genetics / Urinary Bladder Neoplasms - metabolism / Urinary Bladder Neoplasms - pathology / Urothelial cancer / Wound healing / Zoology