Zhang Tao, Mao Changlin, Chang Yao, Lyu Jiaju, Zhao Delong, Ding Sentai
Department of Urology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, China.
Department of Urology, Mindong Hospital Affiliated to Fujian Medical University, Fuan, China.
Curr Urol. 2024 Sep;18(3):185-193. doi: 10.1097/CU9.0000000000000233. Epub 2024 Sep 20.
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.
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.
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.
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.
良性前列腺增生（BPH）的发展与前列腺基质中的缺氧密切相关，并且缺氧诱导因子-1α/血管内皮生长因子（HIF-1α/VEGF）通路已被证明在缺氧反应中会显著激活。该通路在BPH发病机制中激活的潜在机制仍不清楚。
我们构建了HIF-1α过表达和敲低的BPH基质（WPMY-1）和上皮（BPH-1）细胞系，在不同氧气条件（缺氧、常氧和缺氧+HIF-1α抑制剂）下培养。应用定量实时聚合酶链反应（qPCR）和蛋白质印迹法检测HIF-1α/VEGF通路的表达。通过细胞计数试剂盒-8和流式细胞术分析细胞增殖和凋亡。我们使用miRWalk 2.0数据库和蛋白质印迹法预测选择性靶向HIF-1α/VEGF通路的潜在miRNA，并通过qPCR和双荧光素酶测定法验证该预测。
在BPH基质细胞系（WPMY-1）中，VEGF的表达与HIF-1α水平一致，在过表达细胞中升高，在敲低细胞中降低。缺氧诱导HIF-1α过表达，这可被HIF-1α抑制剂逆转。此外，通过细胞计数试剂盒-8和流式细胞术评估，HIF-1α抑制剂在缺氧条件下显著抑制细胞增殖并促进凋亡。然而，在BPH上皮细胞系（BPH-1）中，HIF-1α的表达水平不影响VEGF的表达。最后，从miRWalk 2.0数据库和蛋白质印迹法预测到一种调节HIF-1α/VEGF通路的潜在miRNA，即miR-17-5p，并通过qPCR和双荧光素酶测定法进行了验证。
在缺氧状态下，HIF-1α/VEGF通路的激活在调节BPH基质细胞系中的细胞增殖中起关键作用。miR-17-5p的调节可能是该通路激活的潜在机制。该通路的调节可能参与了BPH的发病机制。
