2001 年度実績報告書

マウス大腸癌転移モデルに基づくヒト大腸癌転移の遺伝子診断法の研究

研究課題

研究課題/領域番号	12217025
研究機関	東京大学
研究代表者	入村達郎東京大学, 大学院・薬学系研究科, 教授 (80092146)
研究分担者	傳田香里東京大学, 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)
キーワード	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
研究概要	癌特定:マウス大腸癌転移モデルに基づくヒト大腸癌の遺伝子診断法の研究宿主との相互作用を考えるために有用である同系マウスを用いた大腸癌肝転移モデルを作成し、肝臓に高転移性のバリアント細胞(colon 38-SL4細胞)を選別し、それ由来のクローン細胞を得た。これらの細胞に特徴的な形質は、肺転移性の高い細胞の形質とは異なり、増殖性が高い一方、運動性、接着性、マトリクス分解酵素のレベルに関しては、親株と大きな差異がなかった。ディファレンシャルディスプレイ法、アレイ解析法、及びサブトラクティブPCR法によって、この高転移バリアント細胞で発現量の変化している複数の遺伝子を同定した。発現レベルの差異は、ノザン解析及び半定量RT-PCR法によって確認した。これらの遺伝子は、これまで肺に高転移性の癌細胞の解析から転移に関係するとされて来た細胞形質とは全く異なる分子をコードしているものであった。新たにサブトラクティブPCR法を用いて同定したSL4細胞に高発現している遺伝子を7つ(Enolase 1、Laminin receptor 1、ATP synthase、Desmin、Tubulin alpha 6、EST : BC008110、及びUba52)昨年度に同定したcolon38細胞に高発現のものとしてNuk/EphB2とPN-1の2種類とSL4細胞に高発現のReelinについてヒト相同遺伝子を同定し競合的なPCR法によるmRNA定量法を確立した。大腸癌切除標本におけるこれらの遺伝子の発現を定量し、臨床病理学的な所見との相関を検討した。Nuk/EphB2の発現が肝転移のある標本で有意に低く、Laminin receptor 1の発現がリンパ節転移のある標本で有意に高いことが明らかになった。

研究成果

(22件)

すべてその他

すべて文献書誌 (22件)

[文献書誌] Kato K.et al.: "Distinct Orders of GaINAc Incorporation into a Peptide with Consecutive Threonines"Biochem Biophys Res Commun.. 287. 110-115 (2001)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] Yim M.et al.: "Mutated plant lectin library useful to identify different cells"Proc Nat Acad Sci USA. 98. 2222-2225 (2001)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] Denda-Nagal K. et al.: "MUC1 in carcinoma-host interactions"Giycoconjugate J. 17. 649-658 (2000)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] Koike J et al.: "Density-dependent induction of TNF-a release from human monocytes by immobilized P-selectin"FEES Lett. 477. 84-88 (2000)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] Nagata K. et al.: "Redistribution of selectin counter-ligands induced by cytokines"Int Immunol.. 12. 487-492 (2000)
[文献書誌] 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)

2001 年度 実績報告書

マウス大腸癌転移モデルに基づくヒト大腸癌転移の遺伝子診断法の研究

研究代表者

入村 達郎 東京大学, 大学院・薬学系研究科, 教授 (80092146)

研究成果

[文献書誌] Kato K.et al.: "Distinct Orders of GaINAc Incorporation into a Peptide with Consecutive Threonines"Biochem Biophys Res Commun.. 287. 110-115 (2001)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] Yim M.et al.: "Mutated plant lectin library useful to identify different cells"Proc Nat Acad Sci USA. 98. 2222-2225 (2001)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] Denda-Nagal K. et al.: "MUC1 in carcinoma-host interactions"Giycoconjugate J. 17. 649-658 (2000)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] Koike J et al.: "Density-dependent induction of TNF-a release from human monocytes by immobilized P-selectin"FEES Lett. 477. 84-88 (2000)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] Nagata K. et al.: "Redistribution of selectin counter-ligands induced by cytokines"Int Immunol.. 12. 487-492 (2000)

[文献書誌] 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)

2001 年度実績報告書

入村達郎東京大学, 大学院・薬学系研究科, 教授 (80092146)