Hypoxia activates the hypoxia-inducible factor-1α/vascular endothelial growth factor pathway in a prostatic stromal cell line: A mechanism for the pathogenesis of benign prostatic hyperplasia

Abstract

Background: The development of benign prostatic hyperplasia (BPH) is closely related to hypoxia in the prostatic stroma, and the hypoxia-inducible factor-1α/vascular endothelial growth factor (HIF-1α/VEGF) pathway has been shown to significantly activate in response to hypoxia. The underlying mechanism for activation of this pathway in the pathogenesis of BPH remains unclear.

Materials and methods: We constructed HIF-1α overexpression and knockdown BPH stromal (WPMY-1) and epithelial (BPH-1) cell lines, which were cultured under different oxygen conditions (hypoxia, normoxia, and hypoxia + HIF-1α inhibitor). Quantitative real-time polymerase chain reaction (qPCR) and Western blotting were applied to detect the expression of the HIF-1α/VEGF pathway. Cell proliferation and apoptosis were analyzed by Cell Counting Kit-8 and flow cytometry. We used the miRWalk 2.0 database and Western blotting to predict the potential miRNA that selectively targets the HIF-1α/VEGF pathway, and verified the prediction by qPCR and dual-luciferase assays.

Results: In a BPH stromal cell line (WPMY-1), the expression of VEGF was in accordance with HIF-1α levels, elevated in the overexpression cells and decreased in the knockdown cells. Hypoxia-induced HIF-1α overexpression, which could be reversed by a HIF-1α inhibitor. Moreover, the HIF-1α inhibitor significantly depressed cellular proliferation and promoted apoptosis in hypoxic conditions, assessed by Cell Counting Kit-8 and flow cytometry. However, in the BPH epithelial cell line (BPH-1), the expression level of HIF-1α did not influence the expression of VEGF. Finally, a potential miRNA, miR-17-5p, regulating the HIF-1α/VEGF pathway was predicted from the miRWalk 2.0 database and Western blotting, and verified by qPCR and dual-luciferase assay.

Conclusions: In hypoxia, activation of the HIF-1α/VEGF pathway plays a crucial role in regulating cell proliferation in a BPH stromal cell line. Regulation by miR-17-5p may be the potential mechanism for the activation of this pathway. Regulation of this pathway may be involved in the pathogenesis of BPH.

Keywords

Benign prostatic hyperplasia / Hypoxia-inducible factor-1α / Vascular endothelial growth factor / Pathogenesis / miR-17-5p

Author summay

TZ and CM contributed equally to this work.

Acknowledgments

None.

Statement of ethics

Neither institutional review board's approval nor participants' consent was applicable for this study.

Conflict of interest statement

JL is an associate editor of Current Urology. SD is an editorial board member of Current Urology. This article was accepted after normal external review. The other authors declare that they have no competing financial interests or personal relationships that influence the work reported in this article.

Funding source

We gratefully acknowledge the financial support granted from the Shandong Province Key Research and Development Projects (no. 2016GSF201147) and the Science and Technology Development Program of Jinan (no. 201704127).

Author contributions

TZ: Methodology, writing - reviewing and editing, software;

CM: Data curation, writing - original draft preparation;

YC: Visualization, investigation;

JL: Supervision;

DZ: Software, validation;

SD: Conceptualization, funding acquisition.

Data availability

The datasets generated during and/or analyzed during the current study are not publicly available, but are available from the corresponding author on reasonable request.

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