miR-148a-3p inhibits the proliferation and migration of bladder cancer via regulating the expression of ROCK-1

Chao Xu; Guanwen Zhou; Zhuang Sun; Zhaocun Zhang; Haifeng Zhao; Xianzhou Jiang

doi:10.7717/peerj.12724

miR-148a-3p inhibits the proliferation and migration of bladder cancer via regulating the expression of ROCK-1

PeerJ. 2022 Jan 26:10:e12724. doi: 10.7717/peerj.12724. eCollection 2022.

Authors

Chao Xu^#^{1

2}, Guanwen Zhou^#¹, Zhuang Sun¹, Zhaocun Zhang¹, Haifeng Zhao¹, Xianzhou Jiang¹

Affiliations

¹ Department of Urology, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China.
² Center for Reproductive Medicine, Cheeloo College of Medicine, Shandong University, Jinan, China.

^# Contributed equally.

Abstract

Purpose: To investigate the mechanism of miR-148a-3p regulating the proliferation and migration of bladder tumor cells.

Materials and methods: We conducted a preliminary study to detect the relative expression of miR-148a-3p in bladder cancer and para-cancerous tissue samples. Three bladder tumor cell lines, T24, 5,637 and UM-UC-3, were selected. The expression levels of miR-148a-3p were artificially regulated with miR-148a-3p mimics and the miR-148a-3p inhibitor. The relative expression levels of miR-148a-3p in the samples of each cell line were determined. Cell Counting Kit-8 (CCK-8) was used to detect cell proliferation, while the effect of the miR-148a-3p mimics and inhibitor on tumor cell migration was detected by wound healing assay. Flow cytometry assay was carried out to explore the effect of miR-148a-3p on cell apoptosis. Dual-luciferase reporter assay was performed in order to verify miR-148a-3p's target gene. The expressions of ROCK-1 and Bcl-2 were analyzed by western blot.

Results: The relative expression of miR-148a-3p in tumor and adjacent tissues was assessed with qRT-PCR (P < 0.05) and found to be significantly lower in the tumor tissues than the adjacent tissues. The data obtained from the CCK-8 and wound healing assay showed that intracellular transfection of miR-148a-3p mimics could inhibit cell proliferation and migration, while the miR-148a-3p inhibitor promoted them. Overexpression of miR-148a-3p promoted cell apoptosis in the T24 and 5,637 cell lines. The dual-luciferase reporter assay verified that ROCK-1 is a direct target of miR-148a-3p. Western blot showed that miR-148a-3p overexpression downregulated the expression of ROCK-1 and Bcl-2, while miR-148a-3p knockdown upregulated the expression of ROCK-1 and Bcl-2.

Conclusions: We confirmed that miR-148a-3p was significantly decreased in bladder cancer cells. miR-148a-3p overexpression inhibited bladder cancer cell proliferation and migration, whereas miR-148a-3p knockdown promoted bladder cancer cell proliferation and migration. Moreover, we found that ROCK-1 was a downstream target of miR-148a-3p. We also found that miR-148a-3p induced cell apoptosis by regulating the expression of Bcl-2. However, the deeper mechanism of this regulatory relationship needs further study.

Keywords: Biological behavior; Bladder cancer; miR-148a-3p; miRNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Movement / genetics
Cell Proliferation / genetics
Humans
MicroRNAs* / genetics
Receptor Protein-Tyrosine Kinases
Urinary Bladder Neoplasms* / genetics
rho-Associated Kinases

Substances

MicroRNAs
Receptor Protein-Tyrosine Kinases
rho-Associated Kinases

Grants and funding

This work was supported by the Key Research and Development Plan of Shandong Province (2017GSF221005) and Jinan Science and Technology Plan (201805036). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.