Grupo de Investigación Cánceres de Origen Epitelial, Área de Bioquímica y Biología Molecular, Departamento de Biología de Sistemas, Facultad de Medicina y Ciencias de la Salud, Universidad de Alcalá, Alcalá de Henares, Madrid, Spain.
Endocrine, Polypeptide and Cancer Institute, Veterans Affairs Medical Center, Miami, Florida, USA.
Prostate. 2022 Jun;82(8):933-941. doi: 10.1002/pros.24339. Epub 2022 Mar 24.
Growth hormone-releasing hormone (GHRH) and its receptors have been implicated in the progression of various tumors. In this study, we analyzed the carcinogenetic potential of exposure to GHRH of a nontumor human prostate epithelial cell line (RWPE-1) as well as its transforming effect in a xenograft model.
We performed cell viability, cell proliferation, adhesion and migration assays. In addition, metalloprotease (MMP)-2 activity by means gelatin zymography, GHRH-R subcellular location using confocal immunofluorescence microscopy and vascular endothelial growth factor (VEGF) levels by enzyme-linked immunoassay were assessed. Besides, we developed an in vivo model in order vivo model to determine the role of GHRH on tumorigenic transformation of RWPE-1 cells.
In cell cultures, we observed development of a migratory phenotype consistent with the gelatinolytic activity of MMP-2, expression of VEGF, as well as E-cadherin-mediated cell-cell adhesion and increased cell motility. Treatment with 0.1 µM GHRH for 24 h significantly increased cell viability and cell proliferation. Similar effects of GHRH were seen in RWPE-1 tumors developed by subcutaneous injection of GHRH-treated cells in athymic nude mice, 49 days after inoculation.
Thus, GHRH appears to act as a cytokine in the transformation of RWPE-1 cells by mechanisms that likely involve epithelial-mesenchymal transition, thus reinforcing the role of GHRH in tumorigenesis of prostate.
生长激素释放激素（GHRH）及其受体与各种肿瘤的进展有关。在这项研究中，我们分析了暴露于非肿瘤人前列腺上皮细胞系（RWPE-1）的 GHRH 对肿瘤发生的潜在影响及其在异种移植模型中的转化作用。
我们进行了细胞活力、细胞增殖、粘附和迁移测定。此外，通过明胶酶谱法评估了金属蛋白酶（MMP）-2 的活性、使用共聚焦免疫荧光显微镜评估了 GHRH-R 的亚细胞定位以及通过酶联免疫吸附试验评估了血管内皮生长因子（VEGF）水平。此外，我们还开发了一种体内模型，以确定 GHRH 在 RWPE-1 细胞致瘤转化中的作用。
在细胞培养中，我们观察到一种迁移表型的发展，与 MMP-2 的明胶酶活性、VEGF 的表达以及 E-钙粘蛋白介导的细胞-细胞粘附以及增加的细胞迁移一致。用 0.1µM GHRH 处理 24 小时可显著增加细胞活力和细胞增殖。在接种后 49 天通过皮下注射 GHRH 处理的细胞在无胸腺裸鼠中发展的 RWPE-1 肿瘤中也观察到 GHRH 的类似作用。
因此，GHRH 似乎作为一种细胞因子在 RWPE-1 细胞的转化中起作用，其机制可能涉及上皮-间充质转化，从而增强了 GHRH 在前列腺肿瘤发生中的作用。
