HGF regulates VEGF expression via the c-Met receptor downstream pathways, PI3K/Akt, MAPK and STAT3, in CT26 murine cells

Matsumura,Atsushi; Kubota,Takeshi; Taiyoh,Hiroaki; Fujiwara,Hitoshi; Okamoto,Kazuma; Ichikawa,Daisuke; Shiozaki,Atsushi; Komatsu,Shuhei; Nakanishi,Masayoshi; Kuriu,Yoshiaki; Murayama,Yasutoshi; Ikoma,Hisashi; Ochiai,Toshiya; Kokuba,Yukihito; Nakamura,Takahiro; Matsumoto,Kunio; Otsuji,Eigo

doi:10.3892/ijo.2012.1728

HGF regulates VEGF expression via the c-Met receptor downstream pathways, PI3K/Akt, MAPK and STAT3, in CT26 murine cells

Authors:
- Atsushi Matsumura
- Takeshi Kubota
- Hiroaki Taiyoh
- Hitoshi Fujiwara
- Kazuma Okamoto
- Daisuke Ichikawa
- Atsushi Shiozaki
- Shuhei Komatsu
- Masayoshi Nakanishi
- Yoshiaki Kuriu
- Yasutoshi Murayama
- Hisashi Ikoma
- Toshiya Ochiai
- Yukihito Kokuba
- Takahiro Nakamura
- Kunio Matsumoto
- Eigo Otsuji
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Affiliations: Division of Digestive Surgery, Department of Surgery, Kyoto Prefectural University of Medicine, Kamigyo-ku, Kyoto 602-8566, Japan, Division of Tumor Dynamics and Regulation, Kanazawa University Cancer Research Institute, Kakuma-machi, Ishikawa 920-1192, Japan
Published online on: December 6, 2012 https://doi.org/10.3892/ijo.2012.1728
Pages: 535-542

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Abstract

In the present study, we assessed the involvement of hepatocyte growth factor (HGF)/c-Met signalling with vascular endothelial cell growth factor (VEGF) and hypoxia inducible factor (HIF)-1α expression in the downstream pathways phosphatidylinositol 3-kinase (PI3K)/Akt, mitogen-activated protein kinase (MAPK) and signal transducer and activator of transcription 3 (STAT3) in CT26 cells, to determine the mechanisms of the potent anti-angiogenic effect of NK4. We established genetically modified CT26 cells to produce NK4 (CT26-NK4). VEGF expression in subcutaneous CT26 tumours in vivo and in culture supernatants in vitro was determined by ELISA. HIF-1α expression in nuclear extracts was evaluated by western blot analysis. VEGF and HIF-1α mRNA levels were examined by real-time reverse transcription-polymerase chain reaction (RT-PCR). The DNA binding activity of HIF-1α was evaluated using an HIF-1α transcription factor assay kit. Our results demonstrated that VEGF expression was reduced in homografts of CT26-NK4 cells, compared to those of the control cells. In vitro, VEGF expression, which was induced by HGF, was inhibited by anti-HGF antibody, NK4 and by kinase inhibitors (PI3K, LY294002; MAPK, PD98059; and STAT3, Stattic). HGF‑induced HIF‑1α transcriptional activity was also inhibited by the kinase inhibitors. Real-time RT-PCR demonstrated that HGF‑induced HIF‑1α mRNA expression was not inhibited by LY294002 and PD98059, but was inhibited by Stattic. These data suggest that the PI3K̸Akt, MAPK and STAT3 pathways, downstream of HGF̸c‑Met signalling, are involved in the regulation of VEGF expression in CT26 cells. HGF̸c‑Met signalling may be a promising target for anti-angiogenic strategies.

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