LGR4 interacts with RhoA to modulate cell survival and epithelial-mesenchymal transition via the Wnt/β-catenin...

Benign prostatic hyperplasia (BPH), a prevalent disorder among elderly men, still has an unclear etiology. Leucine-rich repeat-containing G protein-coupled receptor 4 (LGR4) plays a pivotal role in cellular proliferation, differentiation and epithelial-mesenchymal transition (EMT) in various tissues. This study was designed to investigate the roles of LGR4 in BPH.

Human prostate tissues, cultured human prostate cell (BPH-1 and WPMY-1) were utilized to detect LGR4 expression and localization by immunofluorescence staining, RT-PCR, and Western Blot. LGR4 deficient and overexpression cell models were established. Using these models, the levels of cellular viability, apoptosis, cell cycle were quantified by cell counting kit-8 assay, flow cytometry, respectively. Apoptosis-related proteins, cycle-associated proteins, EMT biomarkers and Wnt/β-catenin pathway were measured by Western Blot. Furthermore, a software, AlphaFold3, was used to perform molecular docking analysis between LGR4 and RhoA. We further studied the interaction between LGR4 and RhoA through Co-Immunoprecipitation assay.

Our research findings demonstrated that LGR4 was overexpressed in BPH and localized within the stromal and epithelial components of prostate. LGR4 deficiency promoted apoptosis, arrested the cell cycle, inhibited cell growth and EMT. Conversely, overexpression of LGR4 completely reversed the alterations induced by LGR4 deficiency. We further showed that LGR4 regulated the Wnt/β-catenin pathway by interacting with RhoA to control cellular proliferation, apoptosis and EMT, while barely affected cell cycle.

Overall, our original data indicate that the upregulated LGR4 could interact with RhoA to exert an inhibitory impact on cell apoptosis while promote cellular proliferation and EMT via Wnt/β-catenin pathway.