1996 年度実績報告書

KITレセプターの突然変異によるマスト細胞腫の発生機構

研究課題

研究課題/領域番号	07670245
研究機関	大阪大学
研究代表者	辻村亨大阪大学, 医学部・附属病院, 助手 (20227408)
研究分担者	北村幸彦大阪大学, 医学部, 教授 (70028520) 岡部勝大阪大学, 微生物研究所・感染動物実験施設, 助教授 (30089875)
キーワード	KITレセプターチロシンキナーゼ / 機能獲得性突然変異 / マスト細胞腫 / トランスジェニックマウス / レトロウイルスベクター
研究概要	c-kitレセプター・チロシンキナーゼ(KIT)には機能獲得性突然変異(キナーゼ領域のアルパラギン酸をバリン或いはチロシンへ置換する点突然変異)が見出されている。インターロイキン3(IL-3)に依存して増殖する培養マスト細胞株IC-2はマスト細胞としての多くの表現型を保つがKITを発現していない。IC-2細胞にアルパラギン酸をバリンへ置換した変異c-kit遺伝子(活性型c-kit遺伝子)をレトロウイルス発現系を用いて導入すると、導入細胞はIL-3やKITのリガンドであるstem cell factor(SCF)を含まない培地でも活発に増殖するようになりヌードマウスに移植すると腫瘍を形成した後白血病化した。また、正常マウスの骨髄細胞に活性型c-kit遺伝子を含むレトロウイルスを感染させると、感染骨髄細胞から造血因子がなくてもメチルセルロースを含む培地で顆粒球マクロファージコロニーばかりでなくマスト細胞コロニーや混合コロニーが出現し、感染骨髄細胞をマウスに移植すると白血病が発症した。主要組織適合抗原クラスIのプロモーターで発現制御される活性型c-kit遺伝子を導入したトランスジェニックマウスでは白血病やリンパ腫が発症した。更に、自律性に増殖する培養マスト細胞腫株FMA3ではKITにアルパラギン酸を他のアミノ酸へ置換する変異を認めなかったが傍細胞膜領域に7個のアミノ酸欠失を認め、この欠失のためにKITがSCFの結合なしに構成的に二量体を形成し活発化することがわかった。この欠失型KITをコードするc-kit遺伝子をレトロウイルス発現系を用いてIL-3依存性IC-2細胞へ導入すると、導入細胞は自律性に増殖するようになりヌードマウスに移植すると腫瘍を形成した後白血病化した。本研究により突然変異により構成的に活性化するKITは生物学的活性をもつことが明らかとなり、活性型KITはマスト細胞の腫瘍化に直接的に関与すると考えられる。

研究成果
(18件)

すべてその他

すべて文献書誌 (18件)

[文献書誌] Hashimoto,K: "Transforming and differentiation-inducing potentials of constitutively activated c-kit mutant genes in the IC-2 murine interleukin-3-dependent mast cell line" Am J Pathol. 148・1. 189-200 (1996)
[文献書誌] Tsujimura,T: "Constitutive activation of c-kit in FMA3 murine mastocytoma cells caused by deletion of seven amino acids at the juxtamembrane domain" Blood. 87・1. 273-283 (1996)
[文献書誌] Moriyama,Y: "Role of aspartic acid 814 in the function and expression of c-kit receptor tyrosine kinase" J Biol Chem. 271・7. 3347-3350 (1996)
[文献書誌] Kitayama,H: "Neoplastic transformation of normal hematopoietic cells by constitutively activating mutations of c-kit receptor tyrosine kinase" Blood. 88・3. 995-1004 (1996)
[文献書誌] Tsujimura,T: "Involvement of transcription factor encoded by the mi locus in the expression of c-kit receptor tyrosine kinase in cultured mast cells of mice" Blood. 88・4. 1225-1233 (1996)
[文献書誌] Morii,E: "Regulation of mouse mast cell protease 6 gene expression by transcription factor encoded by the mi locus" Blood. 88・7. 2488-2494 (1996)
[文献書誌] Kanakura,Y: "The c-kit receptor tyrosine kinase : its roles in hematological malignancies" Cancer J. 9・2. 79-82 (1996)
[文献書誌] Tsujimura,T: "Role of c-kit receptor tyrosine kinase for development,survival and neoplastic transformation of mast cells" Pathol Int. 46・12. 933-938 (1996)
[文献書誌] Adachi,S: "Inhibition of attachment between cultured mast cells and fibroblasts by phorbol 12-myristate 13-acetate and stem cell factor" Exp Hematol. 23・1. 58-65 (1995)
[文献書誌] Kamada,S: "bcl-2 deficiency in mice leads to pleiotropic anbormalities : Accelerated lymphoid cell death in thymus and spleen,polycystic kidney,hair hypopigmentation,and distorted small intestine" Cancer Res. 55・2. 354-359 (1995)
[文献書誌] Kitayama,H: "Constitutive activating mutations of c-kit receptor tyrosine kinase cnfer factor-independent growth and tumorigenicity of factor-dependent hematopoietic cell lines" Blood. 85・3. 790-798 (1995)
[文献書誌] Tsujimura T: "Substitution of an aspartic acid results in constitutive activation of c-kit receptor tyrosine kinase in a rat tumor mast cell line RBL-2H3" Int Arch Allergy Immunol. 106・4. 377-385 (1995)
[文献書誌] Matsumura,I: "Growth response of acute myeloblastic leukemia cells to recombinant human thrombopoietin" Blood. 86・2. 703-709 (1995)
[文献書誌] Kanno,H: "Primary structure of murine red cell-type pyruvate kinase (PK) and molecular characterization of PK deficiency identified in the CBA strain" Blood. 86・8. 3205-3210 (1995)
[文献書誌] Morimoto,M: "Pyruvate kinase deficiency of mice associated with nonpherocytic hemolytic anemia and cure of the anemia by marrow transplantation without host irradiation" Blood. 86・11. 4323-4330 (1995)
[文献書誌] Kitamura,Y: "Regulation of development,survival and neoplastic growth of mast cells through c-kit receptor" Int Arch Allergy Immunol. 107・1〜3. 54-56 (1995)
[文献書誌] Kitamura,Y: "Hematopoietic Lineages : Regulation of cell production and development" marcel dekker,New York, in press
[文献書誌] Kitamura,Y: "Biological and Molecular Aspects of Mast Cell and Basophil Differentiation and Function" Reven Press,New York, 270 (1995)

1996 年度 実績報告書

KITレセプターの突然変異によるマスト細胞腫の発生機構

研究代表者

辻村 亨 大阪大学, 医学部・附属病院, 助手 (20227408)

研究成果

[文献書誌] Hashimoto,K: "Transforming and differentiation-inducing potentials of constitutively activated c-kit mutant genes in the IC-2 murine interleukin-3-dependent mast cell line" Am J Pathol. 148・1. 189-200 (1996)

[文献書誌] Tsujimura,T: "Constitutive activation of c-kit in FMA3 murine mastocytoma cells caused by deletion of seven amino acids at the juxtamembrane domain" Blood. 87・1. 273-283 (1996)

[文献書誌] Moriyama,Y: "Role of aspartic acid 814 in the function and expression of c-kit receptor tyrosine kinase" J Biol Chem. 271・7. 3347-3350 (1996)

[文献書誌] Kitayama,H: "Neoplastic transformation of normal hematopoietic cells by constitutively activating mutations of c-kit receptor tyrosine kinase" Blood. 88・3. 995-1004 (1996)

[文献書誌] Tsujimura,T: "Involvement of transcription factor encoded by the mi locus in the expression of c-kit receptor tyrosine kinase in cultured mast cells of mice" Blood. 88・4. 1225-1233 (1996)

[文献書誌] Morii,E: "Regulation of mouse mast cell protease 6 gene expression by transcription factor encoded by the mi locus" Blood. 88・7. 2488-2494 (1996)

[文献書誌] Kanakura,Y: "The c-kit receptor tyrosine kinase : its roles in hematological malignancies" Cancer J. 9・2. 79-82 (1996)

[文献書誌] Tsujimura,T: "Role of c-kit receptor tyrosine kinase for development,survival and neoplastic transformation of mast cells" Pathol Int. 46・12. 933-938 (1996)

[文献書誌] Adachi,S: "Inhibition of attachment between cultured mast cells and fibroblasts by phorbol 12-myristate 13-acetate and stem cell factor" Exp Hematol. 23・1. 58-65 (1995)

[文献書誌] Kamada,S: "bcl-2 deficiency in mice leads to pleiotropic anbormalities : Accelerated lymphoid cell death in thymus and spleen,polycystic kidney,hair hypopigmentation,and distorted small intestine" Cancer Res. 55・2. 354-359 (1995)

[文献書誌] Kitayama,H: "Constitutive activating mutations of c-kit receptor tyrosine kinase cnfer factor-independent growth and tumorigenicity of factor-dependent hematopoietic cell lines" Blood. 85・3. 790-798 (1995)

[文献書誌] Tsujimura T: "Substitution of an aspartic acid results in constitutive activation of c-kit receptor tyrosine kinase in a rat tumor mast cell line RBL-2H3" Int Arch Allergy Immunol. 106・4. 377-385 (1995)

[文献書誌] Matsumura,I: "Growth response of acute myeloblastic leukemia cells to recombinant human thrombopoietin" Blood. 86・2. 703-709 (1995)

[文献書誌] Kanno,H: "Primary structure of murine red cell-type pyruvate kinase (PK) and molecular characterization of PK deficiency identified in the CBA strain" Blood. 86・8. 3205-3210 (1995)

[文献書誌] Morimoto,M: "Pyruvate kinase deficiency of mice associated with nonpherocytic hemolytic anemia and cure of the anemia by marrow transplantation without host irradiation" Blood. 86・11. 4323-4330 (1995)

[文献書誌] Kitamura,Y: "Regulation of development,survival and neoplastic growth of mast cells through c-kit receptor" Int Arch Allergy Immunol. 107・1〜3. 54-56 (1995)

[文献書誌] Kitamura,Y: "Hematopoietic Lineages : Regulation of cell production and development" marcel dekker,New York, in press

[文献書誌] Kitamura,Y: "Biological and Molecular Aspects of Mast Cell and Basophil Differentiation and Function" Reven Press,New York, 270 (1995)

1996 年度実績報告書

辻村亨大阪大学, 医学部・附属病院, 助手 (20227408)