Medical Research Council Human Reproductive Sciences Unit, The Queen's Medical Research Institute, Little France Crescent, Edinburgh, UK.
Prostate. 2011 Jun 15;71(9):915-28. doi: 10.1002/pros.21308. Epub 2010 Dec 6.
Human metastatic prostate cancer cell growth can be inhibited by GnRH analogs but effects on virus-immortalized prostate cells have not been investigated.
Virus-immortalized prostate cells were stably transfected with rat GnRH receptor cDNA and levels of GnRH binding were correlated with GnRH effects on signaling, cell cycle, growth, exosome production, and apoptosis.
High levels of cell surface GnRH receptor occurred in transfected papillomavirus-immortalized WPE-1-NB26 epithelial cells but not in non-tumourigenic RWPE-1, myoepithelial WPMY-1 cells, or SV40-immortalized PNT1A. Endogenous cell surface GnRH receptor was undetectable in non-transfected cells or cancer cell lines LNCaP, PC3, and DU145. GnRH receptor levels correlated with induction of inositol phosphates, elevation of intracellular Ca(2+) , cytoskeletal actin reorganization, modulation of ERK activation and cell growth-inhibition with GnRH agonists. Hoechst 33342 DNA staining-cell sorting indicated accumulation of cells in G2 following agonist treatment. Release of exosomes from transfected WPE-1-NB26 was unaffected by agonists, unlike induction observed in HEK293([SCL60]) cells. Increased PARP cleavage and apoptotic body production were undetectable during growth-inhibition in WPE-1-NB26 cells, contrasting with HEK293([SCL60]) . EGF receptor activation inhibited GnRH-induced ERK activation in WPE-1-NB26 but growth-inhibition was not rescued by EGF or PKC inhibitor Ro320432. Growth of cells expressing low levels of GnRH receptor was not affected by agonists.
Engineered high-level GnRH receptor activation inhibits growth of a subset of papillomavirus-immortalized prostate cells. Elucidating mechanisms leading to clone-specific differences in cell surface GnRH receptor levels is a valuable next step in developing strategies to exploit prostate cell anti-proliferation using GnRH agonists.
促性腺激素释放激素（GnRH）类似物可以抑制人类转移性前列腺癌细胞的生长，但尚未研究其对病毒永生化前列腺细胞的影响。
病毒永生化前列腺细胞稳定转染大鼠 GnRH 受体 cDNA，并将 GnRH 结合水平与 GnRH 对信号转导、细胞周期、生长、外泌体产生和凋亡的影响相关联。
高表达的细胞表面 GnRH 受体出现在转染的乳头状瘤病毒永生化 WPE-1-NB26 上皮细胞中，但在非肿瘤源性的 RWPE-1、肌上皮 WPMY-1 细胞或 SV40 永生化 PNT1A 细胞中未检测到。内源性细胞表面 GnRH 受体在未转染的细胞或癌症细胞系 LNCaP、PC3 和 DU145 中无法检测到。GnRH 受体水平与诱导肌醇磷酸产生、细胞内 Ca2+ 升高、细胞骨架肌动蛋白重排、ERK 激活的调节以及 GnRH 激动剂诱导的细胞生长抑制相关。Hoechst 33342 DNA 染色-细胞分选表明，在激动剂处理后，细胞在 G2 期积累。与在 HEK293([SCL60])细胞中观察到的诱导作用不同，转染的 WPE-1-NB26 细胞中，激动剂对 exosomes 的释放没有影响。在 WPE-1-NB26 细胞中，生长抑制过程中无法检测到多聚二磷酸腺苷核糖聚合酶（PARP）裂解和凋亡小体的产生，与 HEK293([SCL60]) 形成对比。EGF 受体激活抑制了 WPE-1-NB26 中 GnRH 诱导的 ERK 激活，但 EGF 或 PKC 抑制剂 Ro320432 不能挽救生长抑制。表达低水平 GnRH 受体的细胞生长不受激动剂影响。
工程化高水平 GnRH 受体激活抑制了一组乳头状瘤病毒永生化前列腺细胞的生长。阐明导致细胞表面 GnRH 受体水平克隆特异性差异的机制是下一步开发利用 GnRH 激动剂抑制前列腺细胞增殖的有价值策略。
