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hsa_circ_0000417 downregulation suppresses androgen receptor expression and apoptotic signals in human foreskin fibroblasts via sponging miR-6756-5p

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Abstract

Background

Dysregulated apoptosis of penile mesenchymal cells during male urethragenesis has been previously demonstrated to underly hypospadiac urethral closure failure, and androgen receptor (AR) has been shown to play a central role in regulating penile mesenchyme cell proliferation and survival. However, the regulatory mechanisms upstream and downstream of AR remain poorly understood. Our clinical data and bioinformatics analysis previously indicated that hsa_circ_0000417, a circRNA significantly downregulated in hypospadias preputial specimens, may act as a ceRNA for AR via sequestering hsa_miR-6756-5p, and that the biological functions of hsa_circ_0000417 may significantly involve the PI3K/AKT pathway. In this study, we employed human foreskin fibroblasts (HFF-1) to experimentally validate this putative hsa_circ_0000417/miR-6756-5p/AR axis and its impact on penile mesenchymal cell proliferation and apoptosis.

Method and results

We showed that hsa_circ_0000417 knockdown significantly promoted proliferation and suppressed apoptosis of HFF-1 cells. Mechanistically, hsa_circ_0000417 functioned as a molecular sponge for miR-6756-5p in HFF-1 cells and relieved the latter’s translational repression on AR mRNA, leading to decreased AKT activation and increased expression of pro-apoptotic proteins BAX and cleaved-caspase 9. Conversely, elevated levels of miR-6756-5p resulted in diminished AR expression concomitant with enhanced AKT activation and HFF-1 cell proliferation.

Conclusions

Collectively, our data describe for the first time a circRNA-mediated post-transcriptional regulatory mechanism of AR and its functional consequences in penile mesenchymal cells in the context of hypospadias. These findings may contribute to advancing our current understanding of the roles of AR and mesenchymal cell fate decisions during penile morphogenesis.

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Funding

This work was supported by the National Natural Science Foundation of China (82060129), the “Medical Excellence Award” funded by the Creative Research Development Grant from the First Affiliated Hospital of Guangxi Medical University (2017026), 2018 Guangxi Scholarship Fund of Guangxi Education Department and Key Research and Development Program of Guangxi Province (GuiKe.AB18221028).

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  1. Junqiang Huang and Cheng Su equally contributed to this work and share first authorship.

Authors and Affiliations

  1. Department of Pediatric Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanning, China

    Junqiang Huang, Cheng Su, Xiangyou Zhao, Yuling Liu, Qike Xie & Chao Chen

  2. Faculty of Medicine, Amsterdam UMC, University of Amsterdam, Meibergdreef 9, Amsterdam, Netherlands

    Pingan Lu

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  1. Junqiang Huang
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Contributions

Conceptualization, data curation, investigation, data analysis, visualization: JH, CS, XZ, CC, PL. Software, statistical analysis: YL, QX. Manuscript writing and editing: PL, JH. Supervision: CC.

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Correspondence to Chao Chen.

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Huang, J., Su, C., Lu, P. et al. hsa_circ_0000417 downregulation suppresses androgen receptor expression and apoptotic signals in human foreskin fibroblasts via sponging miR-6756-5p. Mol Biol Rep 50, 6769–6781 (2023). https://doi.org/10.1007/s11033-023-08628-6

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