Testosterone promotes human foreskin fibroblast growth through miR-143-3p targeting IGFBP-3
JOURNAL OF MENS HEALTH(2023)
Abstract
Testosterone is an important male hormone, which could improve the maintenance and recovery of gonadal function in males as well as the repair of human hypospadias and cell fibrosis. Our study focused on investigating the regulatory effect of testosterone in human foreskin fibroblasts (HFF-1) and regulatory mechanisms involved. In this study, HFF-1 cells were treated with testosterone, and cell viability and migration were assessed by cell counting kit-8 (CCK8) and Transwell assays. The expression levels of androgen receptor (AR), miR-143-3p and insulin-like growth factor binding protein-3 (IGFBP3) were measured by quantitative real-time PCR (qRT-PCR), Western blotting, and immunofluorescence. In addition, a potential binding site for miR-143-3p on IGFBP-3 was predicted and its direct binding was further confirmed by a dual luciferase reporter assay. These results showed that testosterone increased the viability and migration of HFF-1 cells. Testosterone could down-regulate miR-143-3p and up-regulate IGFBP3 and AR. Overexpression of miR-143-3p hindered HFF-1 cell viability and negatively regulated IGFBP-3, whereas inhibition of IGFBP-3 impeded cell viability and migration. Furthermore, miR-143-3p was found to directly bind to IGFBP-3. Overexpression of IGFBP-3 countered the regulation of HFF-1 cells by miR-143-3p mimics. In conclusion, this study showed that testosterone promoted the proliferation and migration of HFF1 cells and AR signaling, at least via the miR-143-3p/IGFBP-3 axis. This discovery presents a novel insight for testosterone application in male disorders like hypospadias.
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Key words
Testosterone, miR-143-3p, IGFBP-3, HFF-1 cells
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