がん細胞の薬剤感受性と抵抗性の分子機構とその制御

• Project/Area Number: 06283210
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas (A)
• Allocation Type: Single-year Grants
• Research Institution: The University of Tokyo
• Principal Investigator: 鶴尾 隆 東京大学, 分子細胞生物学研究所, 教授 (00012667)
• Co-Investigator(Kenkyū-buntansha): 藤田 直也 東京大学, 分子細胞生物学研究所, 助手 (20280951) ; 冨田 章弘 (富田 章弘) 東京大学, 分子細胞生物学研究所, 助手 (40251483) ; 清宮 啓之 癌研究会, 癌化学療法センター, 研究員 (50280623) ; 馬島 哲夫 医薬品副作用被害救済, 研究振興調査機構, 派遣研究員 (30311228) ; 内藤 幹彦 東京大学, 分子細胞生物学研究所, 助教授 (00198011) ; 杉本 芳一 東京大学, 癌研究会・癌化学療法センター, 研究員 (10179161)
• Project Period (FY): 1994 – 1999
• Project Status: Completed (Fiscal Year 1999)
• Budget Amount: ¥214,000,000 (Direct Cost: ¥214,000,000); Fiscal Year 1999: ¥40,000,000 (Direct Cost: ¥40,000,000); Fiscal Year 1998: ¥40,000,000 (Direct Cost: ¥40,000,000); Fiscal Year 1997: ¥39,000,000 (Direct Cost: ¥39,000,000); Fiscal Year 1996: ¥35,000,000 (Direct Cost: ¥35,000,000); Fiscal Year 1995: ¥30,000,000 (Direct Cost: ¥30,000,000); Fiscal Year 1994: ¥30,000,000 (Direct Cost: ¥30,000,000)
• Keywords: P-糖タンパク / 抗癌剤耐性 / 耐性タンパク / 耐性遺伝子 / 分子標的 / 耐性克服 / アポトーシス / 癌転移 / 癌移転

Research Abstract

本年度は、がんの本体解明に伴うがん治療の分子標的を意識しつつ以下の5つの課題に取り組んだ。
1.多剤耐性の分子機構と診断・治療
2.ストレス誘導の耐性の分子機構と治療への応用
3.アポトーシス誘導と耐性化の分子機構と治療への応用
4.抗がん剤の新しいターゲットの解明
5.転移がん細胞の特性の解明とその制御
これらの研究目的に対し以下の成果を得ることができた。
1.P糖タンパク阻害剤MS209がMRPを阻害し、MRPによる多剤耐性を克服することを見い出した。またBcl2とp21の発現により癌細胞が多剤耐性を示すことを明らかにした。
2.固形がん内部のストレス環境下でプロテアソーム依存的にトポイソメラーゼIIαの分解が起こることを明らかにし、プロテアソーム阻害剤がトポイソメラーゼII標的抗がん剤の効果を増強することを見い出した。
3.アポトーシスの際に活性化されたCaspase 3により細胞内のActinが切断され、15kDaのActin断片が細胞の形態変化に関与することを示した。新規ヒトIAPファミリーApollonの遺伝子をクローニングし、Apollonが脳腫瘍細胞株で発現していることを明らかにした。
4.新規テロメラーゼ阻害剤としてFJ5002を見い出し、約2μMでテロメラーゼ活性を50%阻害すること、また200nM以下の低濃度でがん細胞にテロメアの短縮・老化を誘導することを明らかにした。
5.血管新生に関わる塩基性FGF(bFGF/FGF-2)の作用機作を検討した結果、bFGFは骨血管内皮細胞からのプロスタグランジンE2産生誘導を介して破骨細胞の形成・分化を誘導していることを明らかにした。またbFGF依存的な破骨細胞形成は、COX-2阻害剤・COX-2 antisense oligoにより阻害されることも明らかにした。

Research Products

• [Publications] Dan, S.: "Activation of c-Abl tyrosine kinase requires caspase activation and is not involved in JNK/SAPK activation during apoptosis of human monocytic leukemia U937 cells"Oncogene. 18. 1277-1283 (1999)
• [Publications] Mashima, T.: "Caspase-mediated cleavage of cytoskeletal actin plays a positive role in the process of morphological apoptosis"Oncogene. 18. 2423-2430 (1999)
• [Publications] Ogiso, Y.: "Glucose starvation and hypoxia induce nuclear accumulation of proteasome in cancer cells"Biochem. Biophys. Res Commun.. 258. 448-452 (1999)
• [Publications] Hirotani, M.: "NH_2-terminal BH4 domain of Bcl-2 is functional for heterodimerization with bax and inhibition of apoptosis"J. Biol. Chem.. 274. 20415-20420 (1999)
• [Publications] Naasani, I.: "FJ5002: A potent telomerase inhibitor identified by exploiting the disease-oriented screening program with COMPARE analysis"Cancer Res.. 59. 4004-4011 (1999)
• [Publications] Yamori, T.: "Potent antitumor activity of MS-247, a novel DNA minor groove binder, evaluated by an in vitro and in vivo human cancer cell line panel"Cancer Res. 59. 4042-4049 (1999)
• [Publications] Zhang,Y.: "Production of interleukin-11 in bone-derived endothelial cells and its role in the forma-tion of osteolytic bone metastasis." Oncogene. 16. 693-703 (1998)
• [Publications] Haga,N.: "2-deoxyglucose inhibits chemotherapeutic drug induced apoptosis in human monocytic leukemia U937 cells with inhibition of c-Jun N-terminal kinase 1/stress-activated protein kinase acitvation." Int.J.Cancer. 76. 86-90 (1998)
• [Publications] Seimiya,H.: "Functional involvement of PTP-U2L in apoptosis subsequent to terminal differentiation of monoblastoid leukemia cells." J.Biol.Chem.273. 21187-21193 (1998)
• [Publications] Dan,S.: "Selective induction of apoptosis in Philadelphia chromosome-positive chronic myelog-enous leukemia cells by an inhibitor of BCR-ABL tyrosine Kinase,CGP 57148." Cell Death and Differ.5. 710-715 (1998)
• [Publications] Fujita,N.: "Acceleration of apoptotic cell death after the cleavage of Bcl-XL protein by caspase-3-like proteases." Oncogene. 17. 1295-1304 (1998)
• [Publications] Motomura,S.: "Inhibition of P-glycoprotein and recovery of drug sensitivity of human acute leukemic blast cells by multidrug resistance (mdr1) antisense oligonucleotides." Blood. 91. 3163-3171 (1998)
• [Publications] Seimiya,H.: "c-Jun NH_2-terminal kinase-mediated activation of interleukin-1β converting enzyme/CED-3-like protease during anticancer drug-induced apoptosis." J.Biol.Chem.272. 4631-4636 (1997)
• [Publications] Naito,M.: "Phosphatidylserine externalization is a downstream event of interleukin-1β-converting enzyme family protease activation during apoptosis." Blood. 89. 2060-2066 (1997)
• [Publications] Mashima,T.: "Actin cleavage by CPP-32/apopain during the development of apoptosis." Oncogene. 14. 1007-1012 (1997)
• [Publications] Torii,S.: "Apoxin I,a novel apoptosis-inducing factor with L-amino acid oxidase activity purified from western diamondback rattelesnake venom." J.Biol.Chem.272. 9539-9542 (1997)
• [Publications] Watanabe,T.: "Regression of established tumors expressing P-glycoprotein by combinations of adriamycin,cyclosporin derivatives,and MRK-16 antibodies." J.Nat'l cancer Inst.89. 512-518 (1997)
• [Publications] Kato,Y.: "Suppression of experimental lung colonization of mouse colon adenocarcinoma 26 in vivo by an anti-idiotype monoclonal antibody recognizing a platelet surface molecule." Cancer Res.57. 3040-3045 (1997)
• [Publications] Ishiguro,T.: "Identification of genes differentially expressed in B16 murine melanoma sublines with different metastatic potentials." Cancer Res.56. 875-879 (1996)
• [Publications] Fujita,N.: "A novel anti-Thy-1 (CD90) monoclonal antibody induces apoptosis in mouse malignant T-lymphoma cells in spite of inducing bcl-2 expression." Int.J.Cancer. 66. 544-550 (1996)
• [Publications] Mikami,K.: "DT-diaphorase as a critical determinant of sensitivity to mitomycin C in human colon and gastric carcinoma cell lines." Cancer Res.56. 2823-2826 (1996)
• [Publications] Naito,M.: "Potentiation of the reversal activity of SDZ PSC833 on multi-drug resistance by an anti-P-glycoprotein monoclonal antibody MRK-16." Int.J.Cancer. 67. 435-440 (1996)
• [Publications] Lee,S.: "Interleukin-7 inhibits apoptosis of mouse malignant T-lymphoma cells by both suppressing the CPP32-like protease activation and inducing the Bcl-2 expression." Oncogene. 13. 2131-2139 (1996)
• [Publications] Tomida,A.: "Glucose-regulated stresses cause decreased expression of cyclin D1 and hypophosphorylation of retinoblastoma protein in human cancer cells." Oncogene. 13. 2699-2705 (1996)
• [Publications] Naito,M.: "Multidrug Resistance in Cancer Cells." John Wiley & Sons Ltd., 507 (1996)
• [Publications] Fujita,N.: "Apoptosis in normal development and cancer." Taylor & Francis Ltd., 304 (1996)
• [Publications] Sato,W.: "Reversal of multidrug resistance by a novel quinoline derivative,MS-209." Cancer Chemother.Pharmacol.35. 271-277 (1995)
• [Publications] Fujita,N.: "Apoptosis inhibition by anti-Mr23,000(Thy-1)monoclonal antibodies without inducing bc1-2expression." Cell Growth & Differentiation. 6. 355-362 (1995)
• [Publications] Seimiya,H.: "Clonig,expression and chromosomal localization of a novel gene for protein tyrosine phosphatase(PTP-U2)induced by various differentiation-inducing agents." Oncogene. 10. 1731-1738 (1995)
• [Publications] Hanaoka,K.: "Involvement of CD45 in adhesion and suppression of apoptosis of mouse malignant T-lymphoma cells." Cancer Res.55. 2186-2190 (1995)
• [Publications] Seimiya,H.: "Phorbol ester-resistant monoblastoid leukemia cells with a functional mitogen-activated protein kinase cascade but without responsive protein tyrosine phosphatases" Oncogene. 11. 2047-2054 (1995)
• [Publications] Mashima,T.: "Identification of actin as a substrate of ICE and ICE-like protease and involvement of an ICE-like protease but not ICE in VP-16-induced U937 apoptosis." Biochem.Biophys.Res.Commun.217. 1185-1192 (1995)
• [Publications] Tsurou,T.: "Anti-Cancer Drugs" Rapid Communications of Oxford Ltd., 213-218 (1995)
• [Publications] Georgopapadakou,N.H.: "Drug transport in antimicrobial and anticancer chemotherapy." Marcel Dekker,Inc., 549-576 (1995)
• [Publications] Gohji,K.: "Regulation of gelatinase production in metastatic renal cell carcinoma by organ-specific fibroblasts." Jpn.J.Cancer Res.85. 152-160 (1994)
• [Publications] Isoai,A.: "Inhibitory effects of tumor invasion-inhibiting factor 2 and its conjugate on disseminating tumor cells." Cancer Res.54. 1264-1270 (1994)
• [Publications] Lee,J-H.: "Nonenzymatic reductive activation of 7-N-{{2-{[2-(g-L-glutamylamino) ethyl] dithio} ethy1}} mitomycin C by thiol molecules:a novel mitomycin C derivative effective on mitomycin C-resistant tumor cells." Cancer Res.54. 2398-2403 (1994)
• [Publications] Park,J-G.: "MDR1 gene expression:Its effect on drug resistance to doxorubicin in human hepatocellular carcinoma cell lines." J.Natl.Cancer Inst.86. 700-705 (1994)
• [Publications] Shirai,A.: "Transport of cyclosporin A across the brain capillary endothelial cell monolayer by P-glycoprotein." Biochim.Biophys.Acta.1222. 400-404 (1994)
• [Publications] Kataoka,S.: "Resistance to antitumor agent-induced apoptosis in a mutant of human myeloid leukemia U937 cells." Exp.Cell Res.215. 199-205 (1994)
• [Publications] 鶴尾 隆: "薬剤耐性の分子機構" 羊土社, 155 (1994)
• [Publications] 鶴尾 隆: "癌転移の分子機構" メジカルビュー社, 174 (1994)