Strategic treatment by means of transduction of gene in Malignant Melanoma

• Project/Area Number: 14370568
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Plastic surgery
• Research Institution: HOKKAIDO UNIVERSITY
• Principal Investigator: SUGIHARA Tsuneki Hokkaido Univ., Hospital, Prof, 医学部・歯学部附属病院, 教授 (20002157)
• Co-Investigator(Kenkyū-buntansha): TSUTSUMIDA Arata Hokkaido Univ., Hospital, Pysician, 医学部・歯学部附属病院, 医員 ; YAMAMOTO Yuhei Hokkaido Univ., Grad School of Medicine, Asso.Prof, 大学院・医学研究科, 助教授 (70271674) ; IGAWA Hiroharu kanagawa Univ., Grad School of Medicine, Prof, 医学部, 教授 (10232159) ; MORIUCHI Tetsuya Hokkaido Univ., Institute for Genetic Medicine, Prof, 遺伝子病制御研究所, 教授 (20174394) ; HAMADA Jun-ichi Hokkaido Univ., Institute for Genetic Medicine, Asso.Prof, 遺伝子病制御研究所, 助教授 (50192703) ; 横山 統一郎 北海道大学, 医学部附属病院, 医員
• Project Period (FY): 2002 – 2003
• Project Status: Completed (Fiscal Year 2003)
• Budget Amount: ¥8,600,000 (Direct Cost: ¥8,600,000); Fiscal Year 2003: ¥1,200,000 (Direct Cost: ¥1,200,000); Fiscal Year 2002: ¥7,400,000 (Direct Cost: ¥7,400,000)
• Keywords: malignant melanoma / keloid-derived fibroblast / keratinocyte / coculture / apoptosis / TGF-β / introduction of gene / 表皮細胞

Research Abstract

We hypothesized that in coculture of Keratinocytes and Malignant Melanoma cells, the apoptosis of melanoma cells would be induced, if TGF-β1 production of keratinocytes by transduction of gene was down-regulated. Firstly we tried to induce apoptosis in melanoma cells by the action of anti-Fas antibody and anti-TGP-β1 antibody. But we failed to induce apoptosis. Therefore we changed to using keloid-derived fibroblasts which is benign tumor instead of melanoma cells.
In this study, we investigated the influence of normal skin-and keloid-derived keratinocytes (NK and KK) on normal skin-and keloid-derived fibroblasts (NF and KF) utilizing a serum-free indirect co-culture system. The keloid-derived fibroblasts showed a greater proliferation and minimal apoptosis when co-cultured with normal skin-or keloid-derived keratinocytes and the results were most significant in the latter. This difference was not observed when the fibroblasts were treated with conditioned media obtained from normal skin-and keloid-derived keratinocytes. However, conditioned media treated groups showed mote proliferation and less apoptosis as compared to the non-conditioned medium treated control groups. We also analyzed the profile of factors involved in cell growth and apoptosis in fibroblasts co-cultured with kelatinocytes. Extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) phosphorylations, expression of Bcl-2 and transforming growth factor-b1 (TGF-b1) were alt significantly up-regulated in the fibroblasts co-cultured with keloid-derived keratinocytes. Together, these results strongly suggest that the overlying keratinocytes of the keloid lesion play an important role in keloidogenesis by promoting more proliferation and less apoptosis in the underlying fibroblasts through paracrine and double paracrine effects

Research Products

• [Publications] Emi Funayama: "Keratinocytes Promote Proliferation and Inhibit Apoptosis of the Underlying Fibroblasts : An Important Role in the Pathogenesis of Keloid"The Journal of Investigative Dermatology. 121(6). 1326-1331 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Emi Funayama.Thinle Chodon, Akihiko Oyama, Tsuneki Sugihara: "Keratinocytes Promote Proliferation and Inhibit Apoptosis of the Underlying Fibroblasts : An Important Role in the Pathogenesis of Keloid"The Journal of Investigative Dermatology. 121(6). 1326-1331 (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Emi Funayama: "Keratinocytes Promote Proliferation and Inhibit Apoptosis of the Underlving Fibroblasts : An Important Role in the Pathogenesis of Keloid"The Society for Investigative Dermatology. 121巻6号. 1326-1331 (2003)