マウスモデルに基づくヒト大腸癌転移形質のバイオインフォーマティックスによる解析

• Project/Area Number: 12217025
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The University of Tokyo
• Principal Investigator: 入村 達郎 東京大学, 大学院・薬学系研究科, 教授 (80092146)
• Co-Investigator(Kenkyū-buntansha): 傳田 香里 (傅田 香里) 東京大学, 大学院・薬学系研究科, 助手 (00313122) ; 築地 信 東京大学, 大学院・薬学系研究科, 助手 (90302611) ; 東 伸昭 東京大学, 大学院・薬学系研究科, 講師 (40302616)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥49,000,000 (Direct Cost: ¥49,000,000); Fiscal Year 2004: ¥10,000,000 (Direct Cost: ¥10,000,000); Fiscal Year 2003: ¥10,100,000 (Direct Cost: ¥10,100,000); Fiscal Year 2002: ¥10,000,000 (Direct Cost: ¥10,000,000); Fiscal Year 2001: ¥9,900,000 (Direct Cost: ¥9,900,000); Fiscal Year 2000: ¥9,000,000 (Direct Cost: ¥9,000,000)
• Keywords: 肝転移モデル / 細胞周期 / 足場非依存的増殖 / 遣伝子発現解析 / p16ink4a / adkn2a / プロテオミックス / レクチンライブラリー / クラスター解析 / 転移モデル / 肝転移 / 足場依存的増殖 / 遺伝子発現解析 / 糖転移酵素 / ムチン / 遺伝子 / 癌 / ゲノム / 生体分子 / 糖鎖 / colon38細胞 / 肝細胞 / 遺伝子発現 / DNAアレイ / ディファレンシャルディスプレイ / RT-PCR / リーリン / 遺伝子診断

Research Abstract

肝臓に対する高転移性を指標にin vivoで5回選別されたマウス大腸癌細胞(SL1〜5細胞)の4回選別後の細胞(SL4)と親株(colon 38)における遺伝子発現の差異を接着条件及び非接着条件下で網羅的に解析した。接着条件下で両細胞の間で発現に大きな差のある遺伝子30個、接着条件と非接着条件を移した時にcolon 38においてのみ発現が変化する遺伝子270個を同定した。17遺伝子は両方に含まれ、この中にCdkn2aが見出された。その発現はcolon 38及びSL1〜2は高いが、SL3〜5では全く見られなかった。遺伝子サイレンシングの原因はDNAメチル化ではなく、遺伝子の欠損による可能性が高い。Cyclin D1の発現レベルがSL1〜5へ転移性と増殖性が高くなるのと併行して上昇した。colon 38とSL4の表面蛋白質の網羅的解析にレクチン結合性を組み合わせ、低転移性細胞にのみ存在するPNA結合性糖蛋白質と高転移性細胞のみに発現するMAH結合性糖蛋白質を複数同定した。これらの差異は、グリコシレーションによることが示唆された。MAHのループCを改変して作製したライブラリーを用いてcolon 38及びSL4を含む大腸癌細胞の表面のプロファイリングを行った。大腸癌細胞は他の細胞と異なるグループとしてクラスターされ、colon 38を含むクラスターとSL4と実験的に転移性が報告されているほとんどのヒト細胞を含むそれが形成された。

Research Products

• [Journal Article] Distribution of MGL1 binding sites and MGL1/2-positive cells in lymphnodes during the sensitization phase of contact hypersensitivity. (2005)
  • Author(s): Sato K, et al.
  • Journal Title: Acta Histochem Cytochem (In press)
• [Journal Article] Redistribution of macrophages expressing the macrophage galactose-type C-type lectin (MGL) during antigen-induced chronic granulation tissue formation. (2005)
  • Author(s): Sato K, et al.
  • Journal Title: Int Immunol (In press)
• [Journal Article] Expression of MUC1 recognized by a monoclonal antibody MY.1E12 is a useful biomarker for tumor aggressiveness of carcnoma f of the gaolbladder or tumor aggressiveness of carcnoma of the gaolbladder. (2004)
  • Author(s): Kawamoto T, et al.
  • Journal Title: Clin Exp Metastasis 21 Pages: 353-362
• [Journal Article] Expression of MUC1 recognized by monoclonal antibody MY.1E12 is a useful biomaker for tumor aggressoveness pf advanced colon carcinoma. (2004)
  • Author(s): Suzuki H, et al.
  • Journal Title: Clin Exp Metastasis 21 Pages: 321-329
• [Journal Article] Identification of sialoadhesin as a dominant lymph node counter-receptor for mouse macrophage galactose-type C-type lectin 1 (2004)
  • Author(s): Kumamoto Y, et al.
  • Journal Title: J Biol Chem 279 Pages: 49274-49280
• [Journal Article] Antitumor activity of siRNA/cationic liposome complex in mouse models of cancer : A new clinical application of si RNA (2004)
  • Author(s): Yano J, et al.
  • Journal Title: Clinical Cancer Res, 10 Pages: 7721-7726
• [Publications] Miyahara N, et al.: "Expression of UDP-N-acetyl-alpha-D-galactosamine-polypeptide N-acetylgalactosaminyltransferase isozyme 3 in the subserosal layer correlates with postsurgical_survival of pathological tumor stage 2 carcinoma of the gallbladder"Clinical Can Res. (In press). (2004)
• [Publications] Hamada T, et al.: "Mucin Expression in Pleomorphic Adenoma of Salivary Gland : A Potential Role of MUC1 as a Marker to Predict a Recurrence"J.Clinical pathology. (In press). (2004)
• [Publications] Tamada Y, et al.: "Model for paraaortic lymph node metastasis produced by orthotopic implantation of ovarian carcinoma cells in athymic nude mice"Euro J.of Can.. 40. 158-163 (2003)
• [Publications] Horinouchi M, et al.: "Expression of Different Glycoforms of Membrane Mucin (MUC1) and Secretory Mucin (MUC2, MUC5AC and MUC6) in Pancreatic Neoplasms"Acta Histochem.Cytochem.. 36(5). 443-453 (2004)
• [Publications] Tsuiji H, et al.: "Aberrant 0-glycosylation inhibits stable expression of dysadherin, a carcinoma-associated antigen, and facilitates cell-cell adhesion"Glycobiology. 13. 521-527 (2003)
• [Publications] Kanoh A, et al.: "Expression levels of FUT6 gene transfected into human colon carcinoma cells switch two sialyl-Lewis X-related carbohydrate antigens with distinct properties in cell adhesion"Biochem biophi res com.. 303. 896-901 (2003)
• [Publications] Higashi N: "The macrophage C-type lectin specific for galactose/N-acetylgalactosamine is endocytic receptor expressed on monocyte-derived immature dendritic cells"J Biol Chem.. 277. 20686-20693 (2002)
• [Publications] Bando T: "Basic studies on a labeled anti-mucin antibody detectable by infrared-fluorescen endoscopy"J Gastroenterol.. 37. 260-269 (2002)
• [Publications] Onami TM: "The generation of mice deficient for the macrophage galactose and acetylgalactosamine specific lectin : limited role in lymphoid and erythroid homeostas and evidence for multiple lectins"Mol Cell Bid.. 22. 5173-5181 (2002)
• [Publications] Tsuji T: "Regulation of melanoma cell migration and invasion by laminin-5 and α 3 βintegrin(VLA-3)"Clin Exp Metastasis. 19. 127-134 (2002)
• [Publications] Denda-Nagai K: "Macrophage C-type lectin (MGL) on bone marrow-derived immature dendritic cells involved in the recognition and internalization of glycosylated antigens"Glycobiology. 12. 443-450 (2002)
• [Publications] Higashi, N: "Human macrophage lectin specific for galactose/N-acetylgalactosamine is a marker cells at an intermediate stage in their differentiation from monocytes into macrophage"Int Immunol. 14. 545-554 (2002)
• [Publications] Tsuiji M: "Molecular cloning and characterization of a novel mouse macrophage C-type lect mMGL2, which has a distinct carbohydrate specificity from mMGL1"J Biol Chem.. 277. 28892-28901 (2002)
• [Publications] Kato T: "Characterization of the primoter for the mouse a3 integrin gene involvement of tets-family of transchption factors in the promoter activity"Eur J Biochem.. 269. 4524-4532 (2002)
• [Publications] Takeuchi H: "The epitope recognized by the unique anti-MUC1 monoclonal antibody MY.1E12 involves sialyl α 2-3galactosyl βl-3N-acetylgalactosaminide linked to a distinct threonine residue in the MUC1 tandem repeat"J Immunol Methods. 270. 199-209 (2002)
• [Publications] Higashi N: "Redistribution of fibroblasts and macropages as micrometastases develop into established liver metastases"Clin Exp Metastasis. 19. 631-638 (2002)
• [Publications] Sugahara D: "Fluorescence Labeling of Oligosaccharides Useful in the Determination of Molecular Interactions"Analytical Sci.. 19. 167-169 (2002)
• [Publications] Takeuchi H: "O-GalNAc incorporation into a cluster acceptor site of three consecutive threonines. Distinct specificity of GalNAc-transferase isoforms"Eur. J. Biochem.. 269. 6173-6183 (2002)
• [Publications] Taki C: "MUC1 mucin expression in follicular dendritic sells and lymphoepithelial lesions of gastric mucosa-associated lymphoid tissue lymphoma"Pathology Int.. 52. 691-701 (2002)
• [Publications] Kamata M: "Vaccination of mice with MUC1 cDNA suppresses the development of lung metastases"Clin Exp metastasis. 19. 689-696 (2002)
• [Publications] Kato K.et al.: "Distinct Orders of GaINAc Incorporation into a Peptide with Consecutive Threonines"Biochem Biophys Res Commun.. 287. 110-115 (2001)
• [Publications] Kato K.et al.: "N-acetylgalactosamine incorporation into a peptide containing consecutive threonine residues by UDP-N-acetyl-D-galactosaminide : polypeptide N-acetylgalactosaminyltransferases"Glycobiology. 11. 812-829 (2001)
• [Publications] Togayachi A. et al.: "Molecular cloning and characterization of UDP-GlcNAc : Lactosylceramide B1,3-N-Acetylgalucosaminyltransferase (B3Gn-T5), an essential enzyme for the expression of HNK-1 and Lewix X epitopes on glycolipids"J Biol Chem.. 276. 22032-22040 (2001)
• [Publications] Yim M.et al.: "Mutated plant lectin library useful to identify different cells"Proc Nat Acad Sci USA. 98. 2222-2225 (2001)
• [Publications] Shirota K. et al.: "Anti-metastatic effect of the sialyl lewis-x analog GSC-1 50 on the human colon carcinoma derived cell line KM12-HX in the mouse"Biol Pharm Bull.. 24. 316-319 (2001)
• [Publications] Kawamoto T, et al.: "Expression of MUC1 mucins with sialylated carbohydrates at the deepest invasding sites correlates with post-surgical prognosis of pT2 carcinoma of the galbladder"Clin Cancer Res. 7. 1333-1342 (2001)
• [Publications] Amano J. et al.: "Intestinal epithelial cells express and secrete the CD43 glycoform that contains core 2 O-glycans"Microbes and Infect. 3. 723-728 (2001)
• [Publications] Uwatoku R. et al.: "Kupffer cell-mediated recruitment of rat dendritic cells to the liver : roles of N-acetylgaractosamine-specific sger recerptors"Gasteroenterology. 121. 1460-1472 (2001)
• [Publications] Morimoto M. et al.: "Fibroblast migratory factor derived from mouse colon carcinoma cells : potential roles of fibronectin in tumor stroma formation"J Cell Biochem.. 80. 635-646 (2001)
• [Publications] Denda-Nagal K. et al.: "MUC1 in carcinoma-host interactions"Giycoconjugate J. 17. 649-658 (2000)
• [Publications] Denda-Nagai K. et al.: "Absence of correlation of MUC1 expression to malignant behavior of renal cell carcinoma in experimental systems"Clin Exp Metastasis. 18. 71-81 (2000)
• [Publications] Kusaka Y. et al.: "Vital immunostaining of human gastric and colorectal cancer grafted into nude mice ; a preclinical assessment of a potential adjunct to videoendoscopy"J Gastroenterol. 35. 748-752 (2000)
• [Publications] Chun K-H. et al.: "Involvement of cytokines in the skin-to-lymph node trafficking of cells of the monocyte-macrophage lineage expressing a C-type lectin"Int immunol.. 12. 1695-1703 (2000)
• [Publications] Ichii S et al.: "Initial steps in lymph node metastasis formation in an experimental system : possible involvement of recognition by macrophage C-type lectins"Cancer Immunol Immunother. 49. 1-9 (2000)
• [Publications] Chun K-H et al.: "Migration of dermal cells expressing a macrophage C-type lectin during the sensitization phase of delayed-type hypersensitivity"J Leukocyte Biol.. 68. 471-478 (2000)
• [Publications] Ota M et al.: "Involvement of cell surface glycans in adhesion of human colon carcinoma cells to liver tissue in a frozen section assay : role of endo b-galactosidase-sensitive structures"Cancer Res.. 60. 5261-5268 (2000)
• [Publications] Koike J et al.: "Density-dependent induction of TNF-a release from human monocytes by immobilized P-selectin"FEES Lett. 477. 84-88 (2000)
• [Publications] Iida S et al.: "Order and maximum incorporation of N-acetyl-D-galactosamine Into threonine residues of MUC2 core peptide with microsome fraction of human-colon-carcinoma-LS174T-cells"Biochem J. 347. 535-542 (2000)
• [Publications] Kang Y et al.: "Increased expression after X-irradiation of MUC1 in cultured human colon carcinoma HT-9 cells"Jpn J Cancer Res.. 91. 324-330 (2000)
• [Publications] Vestergaard C. et al.: "Overproduction of Th2-specific chemokines in NC/Nga mice exhibiting atopi dermatitis-like lesions"J Clin Invest.. 104. 1097-1105 (2000)
• [Publications] Nagata K. et al.: "Redistribution of selectin counter-ligands induced by cytokines"Int Immunol.. 12. 487-492 (2000)
• [Publications] Kimura T. et al.: "Epltope mapping of monoclonal antibodies specific for a macrophage lectin : a calclum-dependent epltope Is In the carbohydrate recognition domain"Mol immunol.. 37. 151-160 (2000)
• [Publications] Nagata,K.: "Redistribution of selectin counter-ligands〜"Int.Immunol.. 12. 487-492 (2000)
• [Publications] Vestergaard,C.: "Over production of Th2-specific chemokires."J.Clinic.Invest.. 104. 1097-1105 (2000)
• [Publications] Kang,Y.: "Increased expression after X-irradiaton〜."Jpn.J.Cancer Res.. 91. 324-330 (2000)
• [Publications] Iida,S.: "Order and maximum incarporation of 〜."Biochem.J.. 347. 535-542 (2000)
• [Publications] Koike,J.: "Density-dependent induction of TNF-a〜."FEBS Lett.. 477. 84-88 (2000)
• [Publications] Ota,M.: "Involvement of cell surface glycans in〜."Cancer Res.. 60. 5261-5268 (2000)
• [Publications] Chun,K.: "Migration of dermal cells expressing 〜."J.Leukocyte Biol.. 68. 471-478 (2000)
• [Publications] Ichii,S.: "Initial steps in lymph node metastasis〜."Cancer Immunol.. 49. 1-9 (2000)
• [Publications] Chun,K.: "Involvement of cytokines in the skin-to〜."Int.Immunol.. 12. 1695-1703 (2000)
• [Publications] Kusaka,Y.: "Vital immunostaining of human gastic〜."J.Gastroenterol.. 35. 748-752 (2000)
• [Publications] Denda-Nagai,K.: "Absence of correlation of Mull〜."Clin.Exp.Metastasis.. 18. 71-81 (2000)
• [Publications] Yim.M: "Mutated plant lectin library useful〜."PNAS. 98. 2222-2225 (2001)
• [Publications] Morimoto,M.: "Fibroblast migratory factor derived from〜."J.Cell.Biochem.. 80. 635-646 (2001)