Effect of heparin-binding EGF-like growth factor and amphiregulin on the MAP kinase-induced production of vascular endothelial growth factor by human granulosa cells

Abstract

The function of granulosa cells is regulated by various hormones and growth factors. Our aim is to clarify the regulation of vascular endothelial growth factor (VEGF) production induced by heparin-binding EGF-like growth factor (HB-EGF) and amphiregulin (AR) in a human granulosa cell line, KGN. KGN cells were cultured and incubated for 24 h with HB-EGF and AR. The levels of VEGF in the culture media were measured by an enzyme-linked immunosorbent assay. The activation of MAP kinase in KGN cells was detected by Western blot analysis. VEGF production was significantly increased by HB-EGF or AR alone in a dose-dependent manner, whereas it was decreased by AG1478 or U0126. The MAP kinase activity was increased by treatment with HB-EGF or AR. The results suggested that VEGF is induced by HB-EGF and AR through mechanisms involving MAP kinase. The increase in VEGF may contribute to neovascularization, which in turn would promote various ovulation phenomena as well as follicular growth.

Pigment epithelium-derived factor regulation by human chorionic gonadotropin in granulosa cells

“…VEGF, a key regulator of ovarian vasculature (Mauro et al 2014) is upregulated by LH/hCG in granulosa cells (Herr et al 2013). The signal transduction that governs hCGinduced VEGF biosynthesis is well documented and is known to be mediated by several factors as PKC (Sriraman et al 2008), PKA (van den Driesche et al 2008) and AREG (Karakida et al 2011)). In this paper, we focus on the signal transduction that governs hCGinduced PEDF downregulation.…”

“…Whereas the signal transduction that governs hCGinduced VEGF biosynthesis is well documented and is known to be mediated by several factors, such as protein kinase C (PKC; Sriraman et al 2008), PKA (van den Driesche et al 2008), and amphiregulin (AREG; Karakida et al 2011), hCG-mediated PEDF downregulation is not yet investigated. In the current study, we report the involvement of PKA and PKC as well as AREG and the downstream effectors ERK1/2 and PI3K, in the hCG-mediated PEDF downregulation.…”

sRAGE downregulates the VEGF expression through EGF-like growth factor signal pathway in PCOS ovarian granulosa cells

“…BTC play a role in VEGF regulation in head and neck squamous carcinoma cells (HNSCCs) [12] . AREG significantly increase VEGF production in the human granulosa cell line, KGN [13] . These results suggest that AREG, BTC, EREG mediate the production of VEGF.…”

The Possible Mechanism of Low-reactive Level Laser Therapy on Infertile Patients

“…血 管 内 皮 増 殖 因 子(vascular endothelial growth factor: VEGF)は 45 kDa の 2 量体糖タンパクであり，血管内皮 細胞の増殖や血管新生，ヒスタミンより強い血管透過性 作用を持つ 20,21) ．排卵前の卵胞液内の VEGF は外層に 拡散し，内莢膜細胞層における血管新生を形成する 22) ． 卵胞液中には，血液中と比較して VEGF をはじめとする 血管新生促進因子が存在し，また，それらの mRNA の 発現が顆粒膜細胞において認められる (Fig.1) 23) ．VEGF は卵胞形成と排卵過程に関わっていると考えられてお り，ART 患者の卵胞液内には高濃度の VEGF は存在し ている 3,4,23) ．卵巣の染色では，発育した卵胞の顆粒膜 細胞に VEGF の発現を認め，卵胞周囲には CD31 で染色 される血管組織を認めた (Fig.2) 24) ．動物実験ではゴナ ドトロピン単独と比較して VEGF の併用により卵胞発 育が促進されることが報告されている 25) ． VEGF は上皮成長因子 (epidermal growth factor: EGF) や EGF 受容体を共有する amphiregulin や heparin-binding EGFlike growth factor (HB-EGF) により産生が増加した．EGF 受 容体の活性化物質を添加すると VEGF 産生は増加するが， EGF 受容体のチロシンキナーゼ阻害剤や MEK 阻害剤を同 時に添加すると VEGF 産生は抑制された (Fig.3) 24) ．次に， VEGF 産生機序における細胞内情報伝達系について検 Fig.2 The localization of VEGF and vesser formation in the ovary. A cell type-specific distribution of VEGF and CD31 expression in normal ovarian tissues was recorded as shown.…”