1993 年度実績報告書

チロシンキナーゼ受容体癌原遺伝子の発現を制御するmi遺伝子の分離同定

研究課題

研究課題/領域番号	05152071
研究機関	大阪大学
研究代表者	野村慎太郎大阪大学, 医学部, 助教授 (80159087)
研究分担者	北村幸彦大阪大学, 医学部, 教授 (70028520) 廣田誠一大阪大学, 医学部, 助手 (50218856) 辻村亨大阪大学, 医学部, 助手 (20227408) 春日井務大阪大学, 医学部, 助手 (80214310)
キーワード	チロシンキナーゼ / 細胞接着 / 骨大理石症マウス / DNA結合タンパク / サブトラクション / マスト細胞 / マウスミュータント / ★骨細胞
研究概要	mi遺伝子に異常があるマウスは骨大理石症、小眼症、メラノサイト欠損症等重篤な症状を示す。これらの症状のかなり多くの部分はmiミュータントマウスにおいて癌原遺伝子であるレセプターチロシンキナーゼc-kit遺伝子の発現が抑制されていることによるものであることが判明した。我々によってmi遺伝子単離が遂行されているときにmi遺伝子そのものと思われる遺伝子の構造の一部が米国の研究者らによって報告された。この情報をもとに我々はその遺伝子を単離し、その遺伝子発現部位をin situハイブリダイゼーション法を用いて検討した。その結果、c-kit発現細胞とmi遺伝子発現細胞にはかなりの共通性が認められた。そのもっともたるものが肥満細胞(マスト細胞)である。実際にmi遺伝子の欠損のあるマウスにおいてマスト細胞の数の減少が認められ、これがc-kit遺伝子の発現に重篤な影響を与えていることがほぼ明らかとなった。我々はさらにこのmi遺伝子の影響が及んでいる遺伝子群がいかなるものかを同定するためにmi遺伝子に欠損のある培養マスト細胞において低いレベルで発現をしている遺伝子を我々の開発したcDNAサブトラクション法によって分離した。それらの構造を検討した結果、分離されたcDNAクローンの一つがマスト細胞特異的なプロテアーゼP6であることが明らかとなった。この遺伝子がmi遺伝子によって直接影響を受けているのか、あるいはc-kit遺伝子産物を介しているのかについては生物学的な検討より前者であることが予想されていたが、実際、P6遺伝子の5'上流にDNA結合蛋白であるmi遺伝子の結合領域を見いだした。現在、それを生化学的な手法によって証明すると共に、mi遺伝子を特異的に認識する抗体を作成している。

研究成果

(27件)

すべてその他

すべて文献書誌 (27件)

[文献書誌] S.Hirota: "Possible involvement of c-kit receptor and its ligand in increase of mast cells in neurofibroma tissues." Arch.Pathol Lab.Med. 117. 996-999 (1993)
[文献書誌] N.Takemura: "Expression and localization of oxitocin receptor gene in human uterine endometrium in relation to the menstrual cycle" Endocrinology. 132. 1830-1835 (1993)
[文献書誌] T.Tsujimura: "Mast cell number in the skin of heterozygote reflects the molecular nature of c-kit mutation." Blood. 81. 2530-2538 (1993)
[文献書誌] T.Kasugai: "Deficient differentiation of mast cells in the skin of mi/mi mice:Usefulness of in situ hybridization for evaluation of mast cell phenotype." Am.J.Pathol. 143. 1337-1347 (1993)
[文献書誌] T.Hirata: "Stem cell factor induces outgrowth of c-kit-positive neurites from dorsal root ganglia of mouse embryos." Development. 119. 49-56 (1993)
[文献書誌] H.Saeki: "Detection of low copy number of Epstein-Barr virus by in situ hybridization by non-radioisotopic probes prepared by polymerase chain reaction." Diagnotic Molecular Pathology. 2. 108-115 (1993)
[文献書誌] S.Osada: "Predominant expression of nPKCη,a Ca2+ -independent isoform of protein kinase C in Epithelial tissue,in association with epithelial differentiation." Cell,Growth and Differentiation. 4. 167-175 (1993)
[文献書誌] S.Hirota: "Expression of osteopontin mRNA by macrophages in atheroscierotic plaques:A possible association with calcification" Am.J.Pathol. 143. 1003-1008 (1993)
[文献書誌] E.Morii: "Types of neurons that express mRNA of c-kit and its ligand in developing murine cerebellum." Acta Histochem,Cytochem. 26. 359-364 (1993)
[文献書誌] J.Nagoshi: "Expression of bone related genes by spindle-shaped and chondrocyte-like cells that appeared in androgen-dependent SC115 tumors after androgen removal." Lab.Invest.(in press).
[文献書誌] T.Nakase: "Expression of bone morphogenetic protein-4 mRNA in the initial stage of fracture healing." J.of Bone and Mineral Res. 9. 651-659 (1994)
[文献書誌] U.Koshimizu: "Wn mutation of c-kit receptor affects its post-translational processing and extracellular expression." Oncogene. 8. 157-162 (1994)
[文献書誌] K.Hirakawa: "Expression and localization of the mRNA for bone matrix proteins during fracture healing revealed as in situ hybridization." J.of Bone and Mineral Res. (印刷中).
[文献書誌] Y.Nakajima: "Identification of a cell types of odontoma-like clusters in microphthalmic(mi/mi)mice using in situ hybridization." J.of Dental Res.(印刷中).
[文献書誌] S.Hirota: "Expression of bone related genes in ectopically developing bone by non-radioisotopic in situ hybridization." Cell and Tissue Res.(印刷中).
[文献書誌] T.Takemura: "Localization of osteopontin in the otolithic organ of adult rats." Hearing Res. (印刷中).
[文献書誌] E.Morii: "Demonstration of retinal cells expressing messenger RNAs of the c-kit receptor and its ligand." Neuroscience Lett.(印刷中).
[文献書誌] T.Yoshino: "Inverse expression of bcl-2 protein and Fas antigen in lymphoblasts in peripheral lymph nodes and activated blood T and B lymphocytes." Blood. (印刷中).
[文献書誌] T.Kuroki: "Protein kinase C in skin:its activation,down regulation,substrate proteins and family members." Excerpta Medica. (印刷中).
[文献書誌] T.Tsujimura: "Ligand-independent activation of c-kit receptor tyrosine kinase in a murine mastocytoma cell linr P-815 generated by a point mutation." Blood. (印刷中).
[文献書誌] A.Ito: "Developmental expression of Flt3 mRNA in the mouse brain." J.of Mol.Neuroscience. (印刷中).
[文献書誌] N.Huh: "Isolation and characterization of a novel hair follicle-specific gene,hac-1." J.Invest.Dermatology. (印刷中).
[文献書誌] H.Kim: "PKCγ gene expression is delayed in postnatal central nervous system of mi/mi mice." J.of Molecular Neuroscience. (印刷中).
[文献書誌] Y.Kaisho: "Expression of neurotrophin genes in the brain of senescence-accerated mouse(SAM)during postnatal development." Brain Res. (印刷中).
[文献書誌] M.Takemura: "Expression and localization of human oxytocin receptor mRNA and its protein in chorion and decidua during parturition." J.Clin.Invest. (印刷中).
[文献書誌] M.Yamazaki: "c-kit gene is expressed by skin mast cells in embryos but not in puppies of W^<sh>/W^<sh> mice : Age dependent abolishment of c-kit gene expression." Blood. (印刷中).
[文献書誌] Y.Kitamura: "In immunopharmacology of Mast cells and lasbphilis" Academic Press London, 232 (1993)

1993 年度 実績報告書

チロシンキナーゼ受容体癌原遺伝子の発現を制御するmi遺伝子の分離同定

研究代表者

野村 慎太郎 大阪大学, 医学部, 助教授 (80159087)

研究成果

[文献書誌] S.Hirota: "Possible involvement of c-kit receptor and its ligand in increase of mast cells in neurofibroma tissues." Arch.Pathol Lab.Med. 117. 996-999 (1993)

[文献書誌] N.Takemura: "Expression and localization of oxitocin receptor gene in human uterine endometrium in relation to the menstrual cycle" Endocrinology. 132. 1830-1835 (1993)

[文献書誌] T.Tsujimura: "Mast cell number in the skin of heterozygote reflects the molecular nature of c-kit mutation." Blood. 81. 2530-2538 (1993)

[文献書誌] T.Kasugai: "Deficient differentiation of mast cells in the skin of mi/mi mice:Usefulness of in situ hybridization for evaluation of mast cell phenotype." Am.J.Pathol. 143. 1337-1347 (1993)

[文献書誌] T.Hirata: "Stem cell factor induces outgrowth of c-kit-positive neurites from dorsal root ganglia of mouse embryos." Development. 119. 49-56 (1993)

[文献書誌] H.Saeki: "Detection of low copy number of Epstein-Barr virus by in situ hybridization by non-radioisotopic probes prepared by polymerase chain reaction." Diagnotic Molecular Pathology. 2. 108-115 (1993)

[文献書誌] S.Osada: "Predominant expression of nPKCη,a Ca2+ -independent isoform of protein kinase C in Epithelial tissue,in association with epithelial differentiation." Cell,Growth and Differentiation. 4. 167-175 (1993)

[文献書誌] S.Hirota: "Expression of osteopontin mRNA by macrophages in atheroscierotic plaques:A possible association with calcification" Am.J.Pathol. 143. 1003-1008 (1993)

[文献書誌] E.Morii: "Types of neurons that express mRNA of c-kit and its ligand in developing murine cerebellum." Acta Histochem,Cytochem. 26. 359-364 (1993)

[文献書誌] J.Nagoshi: "Expression of bone related genes by spindle-shaped and chondrocyte-like cells that appeared in androgen-dependent SC115 tumors after androgen removal." Lab.Invest.(in press).

[文献書誌] T.Nakase: "Expression of bone morphogenetic protein-4 mRNA in the initial stage of fracture healing." J.of Bone and Mineral Res. 9. 651-659 (1994)

[文献書誌] U.Koshimizu: "Wn mutation of c-kit receptor affects its post-translational processing and extracellular expression." Oncogene. 8. 157-162 (1994)

[文献書誌] K.Hirakawa: "Expression and localization of the mRNA for bone matrix proteins during fracture healing revealed as in situ hybridization." J.of Bone and Mineral Res. (印刷中).

[文献書誌] Y.Nakajima: "Identification of a cell types of odontoma-like clusters in microphthalmic(mi/mi)mice using in situ hybridization." J.of Dental Res.(印刷中).

[文献書誌] S.Hirota: "Expression of bone related genes in ectopically developing bone by non-radioisotopic in situ hybridization." Cell and Tissue Res.(印刷中).

[文献書誌] T.Takemura: "Localization of osteopontin in the otolithic organ of adult rats." Hearing Res. (印刷中).

[文献書誌] E.Morii: "Demonstration of retinal cells expressing messenger RNAs of the c-kit receptor and its ligand." Neuroscience Lett.(印刷中).

[文献書誌] T.Yoshino: "Inverse expression of bcl-2 protein and Fas antigen in lymphoblasts in peripheral lymph nodes and activated blood T and B lymphocytes." Blood. (印刷中).

[文献書誌] T.Kuroki: "Protein kinase C in skin:its activation,down regulation,substrate proteins and family members." Excerpta Medica. (印刷中).

[文献書誌] T.Tsujimura: "Ligand-independent activation of c-kit receptor tyrosine kinase in a murine mastocytoma cell linr P-815 generated by a point mutation." Blood. (印刷中).

[文献書誌] A.Ito: "Developmental expression of Flt3 mRNA in the mouse brain." J.of Mol.Neuroscience. (印刷中).

[文献書誌] N.Huh: "Isolation and characterization of a novel hair follicle-specific gene,hac-1." J.Invest.Dermatology. (印刷中).

[文献書誌] H.Kim: "PKCγ gene expression is delayed in postnatal central nervous system of mi/mi mice." J.of Molecular Neuroscience. (印刷中).

[文献書誌] Y.Kaisho: "Expression of neurotrophin genes in the brain of senescence-accerated mouse(SAM)during postnatal development." Brain Res. (印刷中).

[文献書誌] M.Takemura: "Expression and localization of human oxytocin receptor mRNA and its protein in chorion and decidua during parturition." J.Clin.Invest. (印刷中).

[文献書誌] M.Yamazaki: "c-kit gene is expressed by skin mast cells in embryos but not in puppies of W^<sh>/W^<sh> mice : Age dependent abolishment of c-kit gene expression." Blood. (印刷中).

[文献書誌] Y.Kitamura: "In immunopharmacology of Mast cells and lasbphilis" Academic Press London, 232 (1993)

1993 年度実績報告書

野村慎太郎大阪大学, 医学部, 助教授 (80159087)