消化器癌の転移機構とその治療の研究

• Project/Area Number: 14GS0316
• Research Category: Grant-in-Aid for Creative Scientific Research
• Allocation Type: Single-year Grants
• Research Institution: Kyoto University
• Principal Investigator: 武藤 誠 Kyoto University, 医学研究科, 教授 (70281714)
• Co-Investigator(Kenkyū-buntansha): 大島 正伸 金沢大学, がん研究所, 教授 (40324610) ; 青木 正博 京都大学, 医学研究科, 准教授 (60362464) ; 三好 弘之 京都大学, 医学研究科, 助教 (30362479) ; 北村 剛規 京都大学, 医学研究科, 助教 (10378622) ; 石川 智夫 京都大学, 医学研究科, 助手 (70322162) ; 高久 和明 京都大学, 医学研究科, 助手 (90313121)
• Project Period (FY): 2002 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥560,690,000 (Direct Cost: ¥458,330,000、Indirect Cost: ¥102,360,000); Fiscal Year 2006: ¥110,890,000 (Direct Cost: ¥85,300,000、Indirect Cost: ¥25,590,000); Fiscal Year 2005: ¥110,890,000 (Direct Cost: ¥85,300,000、Indirect Cost: ¥25,590,000); Fiscal Year 2004: ¥110,890,000 (Direct Cost: ¥85,300,000、Indirect Cost: ¥25,590,000); Fiscal Year 2003: ¥110,890,000 (Direct Cost: ¥85,300,000、Indirect Cost: ¥25,590,000); Fiscal Year 2002: ¥117,130,000 (Direct Cost: ¥117,130,000)
• Keywords: 転移 / 浸潤 / 同所移植 / マイクロアレイ / ケモカイン / CXCR3 / CCR1 / CCL9 / トランスジェニックマウス / COX-2 / CDX2 / Wnt / LKB1 / 染色体不安定性 / マクロファージ / プロテオミクス / Cdx2 / mTOR / B16F10 / リンパ管新生 / LYVE-1 / mPGES

Research Abstract

1.昨年度までにケモカイン受容体CXCR3がヒト大腸癌細胞株DLD-1のリンパ節転移を促進することを見出していたが、本年度はCXCR3がケモカインの役割として想定されている癌細胞のリンパ節への遊走ではなく、転移先での腫瘍巣形成を促進することを明らかにした。また、ヒト大腸癌臨床検体の解析から、CXCR3の発現が、リンパ節転移との相関が知られているCXCR4の発現よりも強力な予後不良指標となることを示した。
2.昨年度までに、DNAマイクロアレイを用いてColon26マウス大腸癌細胞の高転移株で発現が変動する遺伝子としてAesを同定し、この遺伝子をColon26細胞に誘導的に発現させると浸潤能、転移能がともに有意に減少することを見出していた。当該年度には、Aes遺伝子を安定発現させた大腸癌細胞をマウス直腸に同所移植すると肝臓や肺への転移が有意に減少することを示した。さらに、ルシフェラーゼレポーターを用いた解析により、AesはNotchシグナルを有意に抑制することが分かった。また、ヒト大腸癌の半数以上の症例の転移巣でAesの発現が低下していることを明らかにし、この遺伝子がヒト大腸癌の転移にも重要な役割を果たしている可能性が示唆されたので、Aesの転移抑制作用をさらに明確にするためにAes遺伝子改変マウスを作出した。
3.転移の初期過程である浸潤機構を解明するため、我々の作出した腸癌自然発症モデルマウス(Apc^<Δ716>Smad4複合変異マウス)を解析した。その結果、このマウスの腺癌ではTGF-βファミリーシグナルの遮断により癌細胞におけるケモカインCCL9の発現が増加すること、その受容体CCR1を持つ未分化骨髄球が血中から浸潤先端部へと集簇することを見出した。また、これら未分化骨髄球は細胞外基質分解酵素MMP9,MMP2を産生すること、Apc^<Δ716>Smad4複合変異マウスにCcr1遺伝子の変異を導入すると未分化骨髄球の集簇が消失し癌細胞の浸潤が抑制されることを明らかにした。さらに、一部のヒト大腸癌で、浸潤先端部にCCR1とMMP9を発現する細胞が存在することを示した。

Research Products

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• [Journal Article] Continuous tooth generation in mouse is induced by activated epithelial Wnt/β-catenin signaling. (2006)
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• [Journal Article] The stabilization of β-catenin leads to impaired primordial germ cell development via aberrant cell cycle progression (2006)
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• [Journal Article] Colonic polyposis caused by mTOR-mediated chromosomal instability in Apc^<+/Δ716> Cdx2^<+/-> compound mutant mice. (2003)
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  • Journal Title: Cancer Research 63巻・16号 Pages: 4872-4877
• [Journal Article] Requirement for tumor suppressor Apc in the morphogenesis of anterior and ventral mouse embryo. (2003)
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• [Journal Article] Phospholipase A2 and apoptosis (2002)
  • Author(s): M.M.Taketo
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• [Journal Article] Apc modulates embryonic stem-cell differentiation by controlling the dosage of β-catenin signaling. (2002)
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• [Journal Article] No effects of Smad2 (Madh2) null mutation on malignant progression of intestinal polyps in Apc^<Δ716> knockout mice. (2002)
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• [Journal Article] Hepatocellular carcinoma caused by loss of Heterozygosity in Lkb1 gene knockout mice. (2002)
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• [Journal Article] Gastrointestinal hamartomatous polyposis in Lkb1 heterozygous knockout mice. (2002)
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• [Journal Article] COX selectivity and animal models of colon cancer. (2002)
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• [Journal Article] Lack of tumorigenesis in the mouse liver after adenovirus-mediated expression of a dominant stable mutant of β-catenin (2002)
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• [Journal Article] Imaging Cyclooxygenase-2(Cox-2)Gene Expression in Living Animals with a Luciferase Knock-in Reporter Gene.
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• [Patent(Industrial Property Rights)] 蛍光性蛋白質の非蛍光性変異体を発現するトランスジェニック非ヒト哺乳動物 (2005)
  • Inventor(s): 武藤誠, 園下将大
  • Industrial Property Rights Holder: 国立大学法人京都大学
  • Industrial Property Number: 2005-202465
  • Filing Date: 2005-07-12
• [Patent(Industrial Property Rights)] CXCR3阻害剤を含有する医薬組成物 (2004)
  • Inventor(s): 武藤 誠, 河田 健二
  • Industrial Property Rights Holder: 国立大学法人京都大学
  • Industrial Property Number: 2004-065612
  • Filing Date: 2004-03-09
• [Patent(Industrial Property Rights)] (仮)蛍光性蛋白質の非蛍光性変異を発現するトランスジェニック非ヒト哺乳動物
  • Inventor(s): 武藤 誠, 園下 将大
  • Industrial Property Rights Holder: 国立大学法人京都大学
• [Publications] K.Aoki: "Colonic polyposis caused by mTOR-mediated chromosomal instability in Apc Δ716Cdx2 compound mutant mice."Nature Genetics. 35巻・4号. 323-330 (2003)
• [Publications] M.Morkel: "Beta-catenin regulates Cripto- and Wnt3- dependent gene expression programs in mouse axis and mesoderm formation."Development. 130巻・25号. 6283-6294 (2003)
• [Publications] T.Higuchi: "A randomized, double-blind, placebo-controlled trial of the effects of rofecoxib, a selective cyclooxygenase-2 inhibitor, on rectal polyps in familial adenomatous polyposis patients."Clinical Cancer Research. 9巻・13号. 4756-4760 (2003)
• [Publications] H.Takeda: "Cooperation of cyclooxygenase 1 and cyclooxygenase 2 in intestinal polyposis."Cancer Research. 63巻・16号. 4872-4877 (2003)
• [Publications] N.Soshnikova: "Genetic interaction between Wnt/beta-catenin and BMP receptor signaling during fomation of the AER and the dorsal-ventral axis in the limb."Genes and Development. 17巻・16号. 1963-1968 (2003)
• [Publications] B.Bierie: "Activation of beta-catenin in prostate epithelium induces hyperplasias and squamous transdifferentiation."Oncogene. 22巻・25号. 3875-3887 (2003)
• [Publications] D.Zechner: "Beta-Catenin signals regulate cell growth and the balance between progenitor cell expansion and differentiation in the nervous system."Developmental Biology. 258巻・2号. 406-418 (2003)
• [Publications] T.O.Ishikawa: "Requirement for tumor suppressor Apc in the morphogenesis of anterior and ventral mouse embryo."Developmental Biology. 253巻・2号. 230-246 (2003)
• [Publications] D.Thumkeo: "Targeted disruption of the mouse rho-associated kinase 2 gene results in intrauterine growthretardation and fetal death."Molecular and Cellular Biology. 23巻・14号. 5043-5055 (2003)
• [Publications] K.Miyoshi: "Activation of β-catenin signaling in differentiated mammary secretory cells induces transdifferentiation into epidermis and squamous metaplasias"Proceedings of the National Academy of Sciences or the United States of America. 99巻・1号. 219-224 (2002)
• [Publications] H.Seno: "Cyclooxygenase 2-and prostaglandin E_2 receptor EP_2 -dependent angiogenesis in Apc^<Δ716> mouse intestinal polyps"Cancer Research. 62巻・2号. 506-511 (2002)
• [Publications] N.Harada: "Lack of tumorigenesis in the mouse liver after adenovirus-mediated expression of a dominant stable mutant of β-catenin"Cancer Research. 62巻・7号. 1971-1977 (2002)
• [Publications] M.Oshima: "COX selectivity and animal models for colon cancer"Current Pharmaceutical Design. 8巻・12号. 1021-1034 (2002)
• [Publications] H.Miyoshi: "Gastrointestinal hamartomatous polyposis in Lkbl heterozygous knockout mice"Cancer Research. 62巻・8号. 2261-2266 (2002)
• [Publications] F.Gounari: "Stabilization of β-catenin induces lesions reminiscent of prostatic intraepithelial neoplasia, but terminal squamous transdifferentiation of other secretory epithelia"Oncogene. 21巻・26号. 4099-4107 (2002)
• [Publications] K.Miyoshi: "Activation of different Wnt/ β-catenin signaling components in mammary epithelium induces transdifferentiation and the formation of pilar tumors"Oncogene. 21巻・36号. 5548-5556 (2002)
• [Publications] M.Nakau: "Hepatocellular carcinoma caused by loss of heterozygosity in Lkbl gene knockout mice"Cancer Research. 62巻・16号. 4549-4553 (2002)
• [Publications] K.Takaku: "No effects of Smad2 (Madh2) null mutation on malignant progression of intestinal polyps in Apc^<Δ716> knockout mice"Cancer Research. 62巻・16号. 4558-4561 (2002)
• [Publications] H.Lickert: "Formation of multiple hearts in mice following deletion of β-catenin in the embryonic endoderm"Developmental Cell. 3巻・2号. 171-181 (2002)
• [Publications] H.Seno: "CDX2 expression in the stomach with intestinal metaplasia and intestinal-type cancer : Prognostic implications"International Journal of Oncology. 21巻・4号. 769-774 (2002)
• [Publications] J.Sun: "Hemoprotein Bach 1 regulates enhancer availability of heme oxygenase-1 gene"The EMBO Journal. 21巻・19号. 5216-5224 (2002)
• [Publications] MF.Kielman: "Apc modulates embryonic stem-cell differentiation by controlling the dosage of β-catenin signaling"Nature Genetics. 32巻・4号. 594-605 (2002)
• [Publications] M.Sonoshita: "Cyclooxygenase-2 expression in fibroblasts and endothelial cells of intestinal polyps"Cancer Research. 62巻・23号. 6846-6849 (2002)
• [Publications] M.Matsui: "Mice lacking M2 and M3 muscarinic acetylcholine receptors are devoid of cholinergic smooth muscle contractions but still viable"The Journal of Neuroscience. 22巻・24号. 10627-10632 (2002)
• [Publications] M.Taketo: "Phospolipase A_2 and apoptosis"Biochimica et Biophysica Acta. 1585巻・2-3号. 72-76 (2002)
• [Publications] H.Koizumi: "Targeted disruption of intracellular type I PAF-acetylhydrolase catalytic subunits causes severe impairment in spermatogenesis"The Journal of Biological Chemistry. (発表予定). (2003)
• [Publications] T.Ishikawa: "Requirement for tumor suppressor Apc in the morphogenesis of anterior and ventral mouse embryo"Developmental Biology. 253巻. 230-246 (2003)