2003 年度実績報告書

活性化変異c-Kit,Flt3の腫瘍原性シグナル伝達と標的遺伝子の解析

研究課題

研究課題/領域番号	14570977
研究機関	大阪大学
研究代表者	水木満佐央大阪大学, 医学系研究科, 講師 (80283761)
研究分担者	松村到大阪大学, 医学系研究科, 助教授 (00294083) 金倉譲大阪大学, 医学系研究科, 教授 (20177489)
キーワード	レセプターチロシンキナーゼ / 白血病 / シグナル伝達 / STAT / Pim / 転写因子
研究概要	我々は活性化変異Flt3-ITDの腫瘍特異的シグナル伝達を明らかにするために、骨髄系前駆細胞株32D細胞にFlt3-ITDおよび野生型Flt3を導入した細胞株を作成し、そのmRNAを用いてDNA microchip解析を行った。Flt3-ITD特異的発現誘導遺伝子にはpim-2,SOCS2,SOCS3,CISなどのSTAT3/5標的遺伝子が認められた。Flt3-ITD導入32D細胞は、リガンド刺激Flt3-WTと異なりメチルセルロース上でのコロニー形成能を認めるが、このコロニー形成能はdominant negative STAT5もしくはkinase dead pim-2の導入によって抑制され、Flt3-ITDの腫瘍化においてはSTAT5-Pim-2のシグナル経路が重要な働きを担っていることが示唆された。c-Kit活性化変異体およびFlt3のkinase領域変異体においてもPim-2およびSTAT3/5の標的遺伝子の発現が認められ、STAT3/5-Pim2の経路は活性化変異レセプターチロシンキナーゼ共通の腫瘍化シグナルであることが示唆された。更にPim-2の発現はAML患者由来白血病細胞において、健常骨髄細胞に比較して有意に発現の亢進が認められSTAT3/5-Pim-2の経路はAML腫瘍化に共通のシグナルである可能性が示唆された。Flt3-ITD特異的発現抑制遺伝子には、骨髄系分化に必須の転写因子であるPU.1やC/EBPalphaを認めた。これらの発現は野生型ではリガンド刺激によって誘導され、Flt3-ITDシグナルは野生型シグナルと質的に異なっていた。またFlt3-ITDによって抑制されたC/EBPα、PU.1の発現は、Flt3-ITD特異的キナーゼ阻害剤の処理にて回復し、これらの転写因子の発現抑制がFlt3-ITDシグナル特異的に生じていることが示された。

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] Sonoyama J: "Functional cooperation among Ras, STAT5, and phosphatidylinositol 3-kinase required for full oncogenic activities of BCR/ABL in K562 cells."J Biol Chem.. 277・10. 8076-8082 (2002)
[文献書誌] Nishimura J: "Long-term support of hematopoiesis by a single stem cell clone in patients with paroxysmal nocturnal hemoglobinuria."Blood. 99・8. 2748-2751 (2002)
[文献書誌] Ueda S: "Critical roles of c-Kit tyrosine residues 567 and 719 in stem cell factor-induced chemotaxis : contribution of src family kinase and P13-kinase on calcium mobilization and cell migration"Blood. 99.9. 3342-3349 (2002)
[文献書誌] Tanaka H: "E2F1 and c-Myc potentiate apoptosis through inhibition of NF-kappaB activity that facilitates MnSOD-mediated ROS elimination"Mol Cell. 9・5. 1017-1029 (2002)
[文献書誌] Nanbu-Wakao R: "Stimulation of 3T3-L1 adipogenesis by signal transducer and activator of transcription"Mol Endocrinol. 16・7. 1565-1576 (2002)
[文献書誌] Matsumura I: "Molecular control of megakaryopoiesis and thrombopoiesis"Int J Hematol. 75・5. 473-483 (2002)
[文献書誌] Ezoe S: "GATA-2/estrogen receptor chimera regulates cytokine-dependent growth of hematopoietic cells through accumulation of p21^<WAF1> and p27^<Kip1> proteins."Blood. 100・10. 3512-3520 (2002)
[文献書誌] Ueda S: "Constitutive activation of c-kit by the juxtamembrane but not the catalytic domain mutations is inhibited selectively by tyrosine kinase inhibitors ST1571 and AG1296."Int J Hematol. 76・5. 427-435 (2002)
[文献書誌] Muller-Tidow C: "Mutations of growth factor receptor Flt3 in acute myeloid leukemia : transformation of myeloid cells by Ras-dependent and Ras-independent mechanisms"Dtsch Med Wochenschr. 127・42. 2195-2200 (2002)
[文献書誌] Matsumura I: "Molecular mechanisms of E2F1- and c-Myc-enhanced apoptosis."Recent Res.Devel.Mol.Cell.Biol.. 4. 311-324 (2003)
[文献書誌] Mizuki M: "Constitutive active receptor tyrosine kinase in hematological neoplasia"Recent Res.Devel.Mol.Cell.Biol.. 4. 85-100 (2003)
[文献書誌] Hashimoto K: "Necessity of tyrosine 719 and phosphatidylinositol 3'-kinase-mediated signal pathway in constitutive activation and oncogenic potential of c-kit receptor tyrosine kinase with the Asp814Val mutation"Blood. 101・3. 1094-1102 (2003)
[文献書誌] Mizuki M: "Suppression of myeloid transcription factors and induction of STAT response genes by AML-specific Flt3 mutations"Blood. 101・8. 3164-3173 (2003)
[文献書誌] Zhang X: "Constitutively activated Rho guanosine triphosphatases regulate the growth and morphology of hairy cell leukemia cells"Int.J.Hematol.. 77・3. 263-273 (2003)
[文献書誌] Kawasaki A: "Opposing effects of PML and PML/RARalpha on STAT3 activity"Blood. 101・9. 3668-3673 (2003)
[文献書誌] Matsumura I: "E2FI and c-Myc in cell growth and death"Cell Cycle.. 2・4. 333-338 (2003)
[文献書誌] Mizuki M: "Oncogenic receptor tyrosine kinase in leukemia"Cell Mol Biol.. 49・6. 907-922 (2003)
[文献書誌] Kataoka Y: "Reciprocal inhibition between MyoD and STAT3 in the regulation of growth and differentiation of myoblasts"J Biol Chem.. 278・45. 44178-44187 (2003)

2003 年度 実績報告書

活性化変異c-Kit,Flt3の腫瘍原性シグナル伝達と標的遺伝子の解析

研究代表者

水木 満佐央 大阪大学, 医学系研究科, 講師 (80283761)

研究成果

[文献書誌] Sonoyama J: "Functional cooperation among Ras, STAT5, and phosphatidylinositol 3-kinase required for full oncogenic activities of BCR/ABL in K562 cells."J Biol Chem.. 277・10. 8076-8082 (2002)

[文献書誌] Nishimura J: "Long-term support of hematopoiesis by a single stem cell clone in patients with paroxysmal nocturnal hemoglobinuria."Blood. 99・8. 2748-2751 (2002)

[文献書誌] Ueda S: "Critical roles of c-Kit tyrosine residues 567 and 719 in stem cell factor-induced chemotaxis : contribution of src family kinase and P13-kinase on calcium mobilization and cell migration"Blood. 99.9. 3342-3349 (2002)

[文献書誌] Tanaka H: "E2F1 and c-Myc potentiate apoptosis through inhibition of NF-kappaB activity that facilitates MnSOD-mediated ROS elimination"Mol Cell. 9・5. 1017-1029 (2002)

[文献書誌] Nanbu-Wakao R: "Stimulation of 3T3-L1 adipogenesis by signal transducer and activator of transcription"Mol Endocrinol. 16・7. 1565-1576 (2002)

[文献書誌] Matsumura I: "Molecular control of megakaryopoiesis and thrombopoiesis"Int J Hematol. 75・5. 473-483 (2002)

[文献書誌] Ezoe S: "GATA-2/estrogen receptor chimera regulates cytokine-dependent growth of hematopoietic cells through accumulation of p21^<WAF1> and p27^<Kip1> proteins."Blood. 100・10. 3512-3520 (2002)

[文献書誌] Ueda S: "Constitutive activation of c-kit by the juxtamembrane but not the catalytic domain mutations is inhibited selectively by tyrosine kinase inhibitors ST1571 and AG1296."Int J Hematol. 76・5. 427-435 (2002)

[文献書誌] Muller-Tidow C: "Mutations of growth factor receptor Flt3 in acute myeloid leukemia : transformation of myeloid cells by Ras-dependent and Ras-independent mechanisms"Dtsch Med Wochenschr. 127・42. 2195-2200 (2002)

[文献書誌] Matsumura I: "Molecular mechanisms of E2F1- and c-Myc-enhanced apoptosis."Recent Res.Devel.Mol.Cell.Biol.. 4. 311-324 (2003)

[文献書誌] Mizuki M: "Constitutive active receptor tyrosine kinase in hematological neoplasia"Recent Res.Devel.Mol.Cell.Biol.. 4. 85-100 (2003)

[文献書誌] Hashimoto K: "Necessity of tyrosine 719 and phosphatidylinositol 3'-kinase-mediated signal pathway in constitutive activation and oncogenic potential of c-kit receptor tyrosine kinase with the Asp814Val mutation"Blood. 101・3. 1094-1102 (2003)

[文献書誌] Mizuki M: "Suppression of myeloid transcription factors and induction of STAT response genes by AML-specific Flt3 mutations"Blood. 101・8. 3164-3173 (2003)

[文献書誌] Zhang X: "Constitutively activated Rho guanosine triphosphatases regulate the growth and morphology of hairy cell leukemia cells"Int.J.Hematol.. 77・3. 263-273 (2003)

[文献書誌] Kawasaki A: "Opposing effects of PML and PML/RARalpha on STAT3 activity"Blood. 101・9. 3668-3673 (2003)

[文献書誌] Matsumura I: "E2FI and c-Myc in cell growth and death"Cell Cycle.. 2・4. 333-338 (2003)

[文献書誌] Mizuki M: "Oncogenic receptor tyrosine kinase in leukemia"Cell Mol Biol.. 49・6. 907-922 (2003)

[文献書誌] Kataoka Y: "Reciprocal inhibition between MyoD and STAT3 in the regulation of growth and differentiation of myoblasts"J Biol Chem.. 278・45. 44178-44187 (2003)

2003 年度実績報告書

水木満佐央大阪大学, 医学系研究科, 講師 (80283761)