Role and regulation of phosphatidylinositol 3-kinase in melanoma cells

• Project/Area Number: 17591171
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Dermatology
• Research Institution: Kobe University
• Principal Investigator: OKA Masahiro Kobe University, Kobe University Hospital, Associate Professor, 医学部附属病院, 講師 (30252761)
• Project Period (FY): 2005 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2006: ¥1,700,000 (Direct Cost: ¥1,700,000); Fiscal Year 2005: ¥1,800,000 (Direct Cost: ¥1,800,000)
• Keywords: hepatocyte growth factor / protein kinase C / malignant melanoma / phosphatidylinositol 3-kinase / phosphorylation / Gab 1 / Gab1 / 浸潤 / 転移 / 悪性黒色腫細胞 / アデノウイルスベクター

Research Abstract

The signaling pathway of hepatocyte growth factor (HGF) through its receptor-tyrosine kinase c-Met was examined in human normal melanocytes and three malignant melanoma cell lines MeWo, HM3KO, and HMV-I. HGF-induced c-Met activation was observed in both melanocytes and melanoma cells, whereas phosphatidylinositol 3-kinase (PI3K), a downstream target of c-Met, was not activated in the melanocytes but enhanced in the melanoma cell lines. In contrast, extracellular signal-regulated kinase activity, another target of c-Met, was enhanced by HGF in both the melanocytes and melanoma cells. Therefore, the role of Gab1, the scaffolding adapter protein that couples activated c-Met and PI3K, was analyzed in these cells. Gab1 in the melanocytes showed an electrophoretic mobility slower than that in the melanoma cells, and the mobility shifted to that of the melanoma cells after treatment with alkaline phosphatase, indicating that Gab1 is highly phosphorylated on serine and threonine in the melanocytes. Introduction of protein kinase C (PKC) βII into the melanoma cells, which is expressed in melanocytes but absent in melanoma cells, using an adenovirus vector resulted in serine and threonine phosphorylation of Gab1 and also prevented tyrosine phosphorylation of Gab1 and its association with PI3K. Furthermore, the introduction of PKCβII suppressed HGF-induced activation of PI3K, and attenuated the in vitro invasion activity of the melanoma cells. These results indicate that the HGF signaling process from Gab1 to PI3K is negatively regulated by PKCβII, and that the loss of PKCβII is one of the steps for melanoma cells to gain invasive potential.

Research Products

• [Journal Article] Stimulation of oncogenic metabolic glutamate receptor 1 in melanoma cells activates ERL1/2 via PKCε (2006)
  • Author(s): Marin, Y.E. 他
  • Journal Title: Cell Signal. 18・8 Pages: 1279-1286
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Expression of PKC isoforms in human melanocytic cells in situ. (2006)
  • Author(s): Oka, M. 他
  • Journal Title: J. Dermatol. Sci 41・2 Pages: 157-161
  • NAID: 10020550514
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Overexpression of IL-8 in the cornea induces ulcer formation in the SCID mouse. (2006)
  • Author(s): Oka, M. 他
  • Journal Title: British J. Ophthalmol. 90・5 Pages: 612-615
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Increased expression of germinal center-associated nuclear protein (GANP) is associated with malignant transformation of melanocytes. (2006)
  • Author(s): Kageshita, T. 他
  • Journal Title: J. Dermatol. Sci. 42・2 Pages: 55-63
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Stimulation of oncogenic metabolic glutamate receptor 1 in melanoma cells activates ERK1/2 via PKCε. (2006)
  • Author(s): Marin, Y.E., Namkoong, J., Cohen-Solal, K., Shin, S.S., Martino, J.J., Oka, M., Chen, S.
  • Journal Title: Cell Signal. 18 Pages: 1279-1286
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Expression of PKC isoforms in human melanocytic cells in situ. (2006)
  • Author(s): Oka, M., Kageshita, T., Hsu, M.-Y., Penmatcha, S., Nishigori, C., Herlyn, M.
  • Journal Title: J. Dermatol. Sci. 41 Pages: 157-161
  • NAID: 10020550514
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Overexpression of IL-8 in the cornea induces ulcer formation in the SCID mouse. (2006)
  • Author(s): Oka, M., Norose, K., Matsushima, K., Nishigori, C., Herlyn, M.
  • Journal Title: British J. Ophthalmol. 90 Pages: 612-615
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Increased expression of germinal center-associated nuclear protein (GANP) is associated with malignant transformation of melanocytes. (2006)
  • Author(s): Kageshita, T., Kuwahara, K., Oka, M., Ma, D., Ono, T., Sakaguchi, N.
  • Journal Title: J. Dermatol. Sci. 42 Pages: 55-63
  • NAID: 10020550048
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Stimulation of oncogenic metabolic glutamate receptor 1 in melanoma cells activates ERL1/2 via PKCε. (2006)
  • Author(s): Marin, Y.E., Namkoong, J., Cohen-Solal, K., Shin, S.S., Martino, J.J., Oka, M., Chen, S.
  • Journal Title: Cell Signal. 18・8 Pages: 1279-1286
• [Journal Article] Expression of PKC isoforms in human melanocytic cells in situ. (2006)
  • Author(s): Oka, M., Kageshita, T., Hsu, M.-Y., Penmatcha, S., Nishigori, C., Herlyn, M.
  • Journal Title: J. Dermatol. Sci. 41・2 Pages: 157-161
  • NAID: 10020550514
• [Journal Article] Overexpression of IL-8 in the cornea induces ulcer formation in the SCID mouse. (2006)
  • Author(s): Oka, M., Norose, K., Matsushima, K., Nishigori, C., Herlyn, M.
  • Journal Title: British J. Ophthalmol 90・5 Pages: 612-615
• [Journal Article] Increased expression of germinal center-associated nuclear protein (GANP) is associated with malignant transformation of melanocytes. (2006)
  • Author(s): Kageshita, T., Kuwahara, K., Oka, M., Ma, D., Ono, T., Sakaguchi, N.
  • Journal Title: J. Dermatol. Sci. 42・2 Pages: 55-63
  • NAID: 10020550048
• [Journal Article] Protein kinase C in melanoma. (2005)
  • Author(s): Oka, M.
  • Journal Title: Cancer Metastasis Rev. 24・2 Pages: 287-300