Tekpetey F R, Daniel S A, Yuzpe A
Department of Obstetrics & Gynaecology, University of Western Ontario, London, Canada.
J Assist Reprod Genet. 1995 Nov;12(10):720-7. doi: 10.1007/BF02212900.
The purpose of this study was to investigate the expression of EGF receptor (EGF-R) in human granulosa cells undergoing luteinization and progesterone production by these cells in response to EGF an TGF-alpha alone or in combination with luteinizing hormone (LH). Granulosa cells were obtained from IF patients following oocyte retrieval 34 to 36 hr post-hCG injection. EGF receptor was localized in cells by means of immunoperoxidase staining using a polyclonal primary antibody directed against the human EGF-R. To assess progesterone production, cells were seeded overnight, washed, and cultured with the growth factors +/- LH. Medium and treatments were changed every 24 hr for 3 days.
Specific EGF-R staining was observed in the cultured cells compared to those incubated with antibody that was preabsorbed with a 10-fold excess of EGF. Basal progesterone accumulation per 24-hr period was stimulated dose dependently on each day of culture, by both EGF (up to 3.5-fold at 5 or 50 ng/ml) and TGF-alpha (up to 4-fold at 50 ng/ml). The addition of LH alone also stimulated progesterone accumulation daily, and this effect was further enhanced dose dependently by cotreatment with EGF or TGF-alpha. Furthermore, tyrphostin, an EGF-R-specific tyrosine kinase inhibitor, inhibited both basal and growth factor-stimulated progesterone production.
These data suggest an EGF receptor-mediated physiological role for EGF and TGF-alpha in human luteal function involving an autocrine and/or a paracrine stimulation of progesterone production.
本研究旨在调查表皮生长因子受体（EGF-R）在人黄素化颗粒细胞中的表达，以及这些细胞对单独的表皮生长因子（EGF）和转化生长因子-α（TGF-α）或与促黄体生成素（LH）联合使用时产生孕酮的情况。在注射人绒毛膜促性腺激素（hCG）后34至36小时，从接受体外受精（IVF）的患者中获取颗粒细胞。使用针对人EGF-R的多克隆一抗，通过免疫过氧化物酶染色将EGF受体定位在细胞中。为了评估孕酮的产生，将细胞接种过夜，洗涤后，与生长因子±LH一起培养。每24小时更换一次培养基和处理方式，持续3天。
与用10倍过量的EGF预吸收的抗体孵育的细胞相比，在培养的细胞中观察到特异性EGF-R染色。在培养的每一天，EGF（5或50 ng/ml时高达3.5倍）和TGF-α（50 ng/ml时高达4倍）均以剂量依赖性方式刺激每24小时的基础孕酮积累。单独添加LH也每天刺激孕酮积累，并且与EGF或TGF-α共同处理可使这种作用进一步剂量依赖性增强。此外，EGF-R特异性酪氨酸激酶抑制剂 tyrphostin 抑制基础和生长因子刺激的孕酮产生。
这些数据表明，EGF和TGF-α在人黄体功能中具有EGF受体介导的生理作用，涉及自分泌和/或旁分泌刺激孕酮的产生。
