Qian Qiaofeng, He Weixiang, Liu Daoquan, Yin Jing, Ye Linpeng, Chen Ping, Xu Deqiang, Liu Jianmin, Li Yan, Zeng Guang, Li Mingzhou, Wu Zhonghua, Zhang Yingao, Wang Xinghuan, DiSanto Michael E, Zhang Xinhua
Department of Urology, Zhongnan Hospital of Wuhan University, Wuhan, China.
Department of Rehabilitation, Zhongnan Hospital of Wuhan University, Wuhan, China.
Prostate. 2021 Jun;81(9):530-542. doi: 10.1002/pros.24131. Epub 2021 Apr 16.
Benign prostatic hyperplasia (BPH) is a common disease in elderly men and is often accompanied by chronic inflammation. Macrophages (several subtypes) are the main inflammatory cells that infiltrate the hyperplastic prostate and are found to secrete cytokines and growth factors. The current study aims to explore the effect of M2a macrophages on the development of BPH via insulin-like growth factor 1 (IGF-1).
Human prostate tissues, prostate, and monocyte cell lines (WPMY-1, BPH-1, and THP-1) were used. THP-1 was polarized into several subtypes with cytokines. The expression and localization of IGF-1 and M2 macrophages were determined via immunofluorescent staining, quantitative real-time polymerase chain reaction, and Western blot analysis. Flow cytometry and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays were used to investigate the effects of different subtypes of macrophages on prostate cells. IGF-1 in WPMY-1 and BPH-1 cells was silenced and cocultured with or without M2a macrophages. Cell proliferation, apoptosis, cell cycle, epithelial-mesenchymal transition (EMT), and fibrosis processes were examined.
The polarized subtypes of macrophages were verified by amplifying their specific markers. M2a macrophages enhanced prostate cell proliferation more significantly and CD206 was more expressed in hyperplastic prostate. IGF-1 was localized in both epithelial and stromal components of prostate and upregulated in BPH tissues. M2a macrophages expressed more IGF-1 than other subtypes. Knockdown of IGF-1 in WPMY-1 and BPH-1 cells attenuated cell proliferation, promoted cell apoptosis, retarded cell cycle at the G0/G1 phase, and suppressed the EMT process in BPH-1 cells as well as the fibrotic process in WPMY-1 cells, which was reversible when cocultured with M2a macrophages.
These data demonstrated that knockdown of IGF-1 expression in cultured BPH epithelial and stromal cells reduces proliferation and increases apoptosis. These effects are reversed by coculture with M2a macrophages.
良性前列腺增生（BPH）是老年男性的常见疾病，常伴有慢性炎症。巨噬细胞（几种亚型）是浸润增生前列腺的主要炎症细胞，且发现其可分泌细胞因子和生长因子。本研究旨在通过胰岛素样生长因子1（IGF-1）探讨M2a巨噬细胞对BPH发生发展的影响。
使用人前列腺组织、前列腺及单核细胞系（WPMY-1、BPH-1和THP-1）。用细胞因子将THP-1极化为几种亚型。通过免疫荧光染色、定量实时聚合酶链反应和蛋白质印迹分析确定IGF-1和M2巨噬细胞的表达及定位。采用流式细胞术和3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法研究不同亚型巨噬细胞对前列腺细胞的影响。使WPMY-1和BPH-1细胞中的IGF-1沉默，并与M2a巨噬细胞共培养或不共培养。检测细胞增殖、凋亡、细胞周期、上皮-间质转化（EMT）和纤维化过程。
通过扩增其特异性标志物验证了巨噬细胞的极化亚型。M2a巨噬细胞更显著地增强前列腺细胞增殖，且CD206在增生前列腺中表达更高。IGF-1定位于前列腺的上皮和间质成分中，且在BPH组织中上调。M2a巨噬细胞比其他亚型表达更多的IGF-1。WPMY-1和BPH-1细胞中IGF-1的敲低减弱了细胞增殖，促进了细胞凋亡，使细胞周期阻滞在G0/G1期，并抑制了BPH-1细胞中的EMT过程以及WPMY-1细胞中的纤维化过程，当与M2a巨噬细胞共培养时这是可逆的。
这些数据表明，培养的BPH上皮和间质细胞中IGF-1表达的敲低会降低增殖并增加凋亡。与M2a巨噬细胞共培养可逆转这些效应。
