薬剤感受性と耐性克服の分子生物学的研究

• Project/Area Number: 12217165
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: National Cancer Center Research Institute and Research Center for Innovative Oncology, National Cancer Center Hospital East
• Principal Investigator: 西尾 和人 国立がんセンター(研究所), 薬効試験部, 室長 (10208134)
• Project Period (FY): 2000 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥50,800,000 (Direct Cost: ¥50,800,000); Fiscal Year 2004: ¥10,800,000 (Direct Cost: ¥10,800,000); Fiscal Year 2003: ¥9,800,000 (Direct Cost: ¥9,800,000); Fiscal Year 2002: ¥10,000,000 (Direct Cost: ¥10,000,000); Fiscal Year 2001: ¥10,200,000 (Direct Cost: ¥10,200,000); Fiscal Year 2000: ¥10,000,000 (Direct Cost: ¥10,000,000)
• Keywords: 遺伝子発現解析 / 分子標的薬 / EGFR / 臨床第I相試験 / 血管新生阻害剤 / チロシンキナーゼ阻害剤 / ゲフィチニブ / 臨床付随研究 / 薬理学 / トランスレーショナルリサーチ / マクロアレイ / 薬剤反応性 / シグナル伝達 / 相乗効果 / ZD1839 / ファルネシルトランスフェラーゼ / アポトーシス / Ras / MAPK / cDNAアレイ / 細胞周期制御薬 / ヒートショック / バイオマーカー / 抗がん剤

Research Abstract

各種分子標的薬の抗腫瘍効果規定因子の検討をおこなった。
(1)EGFR特異的チロシンキナーゼ阻害剤ゲフィチニブの感受性はがん細胞のEGFRの発現の有無、自己リン酸化に相関することを示した。
(2)大腸がんおよび肺がんにおけるゲフィチニブとCPT-11の併用の有用性をin vitro, in vivoでしめした。EGFRの各リン酸化欠損株の樹立をおこない、EGFRリン酸化(特にTyr1068)が併用効果のバイオマーカであることを示した。
(3)抗がん剤併用療法におけるcDNAアレイを用いた評価方法の確立を試みた。
(4)細胞周期作用薬E7070に対する耐性細胞を樹立し、同薬剤の感受性マーカーとしてcyclin B2等を選択した。
(5)血管新生阻害薬はVEGFあるいはオステオポンチンの過剰発現細胞による血管新生をより効果的に抑制することを示した。
(6)細胞株レベルで各種がん細胞に薬剤接触し、遺伝子発現の変化をcDNAアレイで検討することで、薬剤特異的なバイオマーカーの選択をおこなった。また同法によって遺伝子発現変動に基づく微小管作用薬の分類をおこなった。
(7)各種臨床検体のRNA抽出、保存、増幅、統計解析等の基礎的な検討をおこなった。統計解析では、再現性評価のためサンプルを測定し級内相関などによりアレイの評価をおこない、日間変動より日内変動が大きいことをしめし、必要なサンプル数が設定できた。また統計解析法、データの標準化の工夫をおこなった。
(8)分子標的薬(各種チロシンキナーゼ阻害剤)の臨床第1相試験における薬物投与前後の患者末梢血単核球における遺伝子発現プロファイルをcDNA遺伝子発現アレイを用いて検討し薬剤が生物学的に適切と考えられる分子に作用している原理の証明が達成できた。有害事象に相関するバイオマーカを選択した。

Research Products

• [Journal Article] Randomized Pharmacokinetic and Pharmacodynamic Study of Docetaxel : Dosing Based on Body-Surface Area Compared With Individualized Dosing Based on Cytochrome P450 Activity Estimated Using a Urinary Metabolite of Exogenous Cortisol. (2004)
  • Author(s): Yamamoto, N.
  • Journal Title: J.Clin.Oncol. 23 Pages: 1-9
• [Journal Article] Small in-frame deletion in the epidermal growth factor receptor as a target for ZD6474. (2004)
  • Author(s): Arao, T.
  • Journal Title: Cancer Res. 64 Pages: 9101-9104
• [Journal Article] Anticancer effects of ZD6474, a VEGF receptor tyrosine kinase inhibitor, in gefitinib ("Iressa")-sensitive and resistant xenog raft models. (2004)
  • Author(s): Taguchi, F.
  • Journal Title: Cancer Sci. 95 Pages: 984-989
• [Journal Article] Synergistic interaction between gefitinib (Iressa, ZD1839) and paclitaxel against human gastric carcinoma cells. (2004)
  • Author(s): Park, J.K.
  • Journal Title: Anticancer Drugs 15 Pages: 809-818
• [Journal Article] The translational study for lung cancer. (2004)
  • Author(s): Nishio, K.
  • Journal Title: Lung Cancer 45
• [Journal Article] hnRNP L enhances sensitivity of the cells to KW-2189. (2004)
  • Author(s): Taguchi, F.
  • Journal Title: Int.J.Cancer 108 Pages: 679-685
• [Journal Article] Synergistic interaction between the EGFR tyrosine kinase inhibitor gefitinib (‘Iressa') and the DNA topoisomerase I inhibitor CPT-11 (Irinotecan) in human colorectal cancer cells. (2004)
  • Author(s): Koizumi, F.
  • Journal Title: Int.J.Cancer 108 Pages: 464-472
• [Journal Article] Development and biological analysis of peritoneal metastasis mouse models for human scirrhous stomach cancer.
  • Author(s): Yanagihara, K.
  • Journal Title: Cancer Sci. (印刷中)
• [Journal Article] Gefitinib treatment affects androgen levels in non-small cell lung cancer patients.
  • Author(s): Nishio, M.
  • Journal Title: Br.J.Cancer (印刷中)
• [Journal Article] The role of DNA-microarray in translational cancer research.
  • Author(s): Korfbe, S.
  • Journal Title: Curr.Pharmacogenomics (印刷中)
• [Journal Article] Establishment of a human non-small cell lung cancer cell line resistant to gefitinib.
  • Author(s): Koizumi, F.
  • Journal Title: Int.J.Cancer (印刷中)
• [Publications] Taguchi, F., Nishio K., et al.: "hnRNP L enhances sensitivity of the cells to KW-2189"Int.J.Cancer. 108. 679-685 (2004)
• [Publications] Koizumi, F., Nishio, K., et al.: "Synergistic interaction between the EGFR tyrosine kinase inhibitor gefitinib (IRESSA') and the DNA topoisomerase I inhibitor CPT-11 (irinotecan) in human colorectal cancer cells."Int.J.Cancer. 108. 464-472 (2004)
• [Publications] Nishiyama, N., Nishio, K., et al.: "Novel cisplatin-incorporated polymeric micelles can eradicate solid tumors in Mice."Cancer Res.. 63. 8977-8983 (2003)
• [Publications] Suzuki, T., Nishio, K., et al.: "MRP5b/SMRP mRNA is highly expressed in metallothionein-deficient mouse liver."J.Health Sci.. 49. 524-526 (2003)
• [Publications] Tsunoda, T., Nishio, K., et al.: "Differential gene expression profiles and identification of the genes relevant to clinicopathologic factors in colorectal cancer selected by cDNA array method in combination with principal component analysis."Int.J.Oncol.. 23. 49-59 (2003)
• [Publications] Usuda, J., Nishio, K., et al.: "Restoration of p53 gene function in 12-O-tetradecanoylphorbor 13-acetate-resistant human leukemia K562/TPA cells."Int.J.Oncol.. 22. 81-86 (2003)
• [Publications] Saijo, N., Nishio, K., et al.: "Translational and clinical studies of target-based cancer therapy."Int.J.Clin.Oncol.. 8. 187-192 (2003)
• [Publications] Kanzawa, F., Nishio, K., et al.: "In vitro effects of combinations of cis-amminedichloro (2-methylpyridine) platinum (II) (ZD0473) with other movel anticancer drugs on the growth of SBC-3, a human small cell lung cancer cell line."Lung Cancer. 40. 325-332 (2003)
• [Publications] Nishiyama, N., Nishio, K., et al.: "Differential gene expression profile between PC-14 cells treated with free cisplatin and cisplatin-incorporated polymeric micelles."Bioconjug.Chem.. 14. 449-457 (2003)
• [Publications] Hirama, M., Nishio, K., et al.: "Osteopontin overproduced by tumor cells acts as a potent angiogenic factor contributing to tumor growth."Cancer Lett.. 198. 107-117 (2003)
• [Publications] Natsume, T., Nishio, K., et al.: "Antitumor activity of TZT-1027 (Soblidotin) against VEGF-secreting human lung cancer in vivo."Cancer Sci. 94. 826-833 (2003)
• [Publications] Saijo, N., Nishio, K., et al.: "Strategy for the development of novel anticancer drugs."Cancer Chemother.Pharmacol.. 52. S97-S101 (2003)
• [Publications] Yamanaka, R., Nishio, K., et al.: "Selection of surrogate marker genes in primary central nervous system lymphomas for radio-chemotherapy by DNA array analysis of gene expression profiles."Int.J.Oncol.. 23. 913-923 (2003)
• [Publications] Kawamura-Akiyama, Y., et al.: "Non-cross resistance of ZD0473 in acquired cisplatin-resistant lung cancer cell lines"Lung Cancer. 38・1. 43-50 (2002)
• [Publications] Naruse, I., et al.: "Enhanced anti-tumor effect of trastuzumab in combination with cisplatin"Jpn.J.Cancer Res.. 93・5. 574-581 (2002)
• [Publications] Fukuoka, K., et al.: "Mechanism of vinorelbine-induced radiosensitization of human small cell lung cancer cells"Cancer Chemother.Pharmacol.. 49・5. 385-390 (2002)
• [Publications] Takahashi, F., et al.: "Osteopontin induces angiogenesis of murine neuroblastoma cells in mice"Int.J.Cancer. 98・5. 707-712 (2002)
• [Publications] Natsume, T., et al.: "Enhanced antitumor activities of TZT-1027 against TNF-α or IL-6 secreting Lewis lung carcinoma in vivo"Cancer Chemother.Pharmacol.. 49・1. 35-47 (2002)
• [Publications] Koh, Y., et al.: "Decreased expression of a2,8 sialyltransferase and increased expression of β1,4 N-acetylgalactosaminyltransferase in gastrointestinal cancers"Proc.Soc.Exp.Biol.Med.. 227・3. 196-200 (2002)
• [Publications] Naruse, I., et al.: "Antitumor activity of the selective epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) Iressa TM (ZD 1839) in a EGFR-expressing multidrug resistant cell line in vitro and in vivo"Int.J.Cancer. 98・2. 310-315 (2002)
• [Publications] Ohira, T., et al.: "Up-regulated gene expression of angiogenesis factors in post-chemotherapeutic lung cancer tissues determined by cDNA macroarray"Oncol.Rep.. 9・4. 723-728 (2002)
• [Publications] Usuda, J., et al.: "Related articles, links restoration of p53 gene function in 12-O-tetradecanoylphorbor 13-acetate-resistant human leukemia K562/TPA cells"Int.J.Oncol.. 22・1. 81-86 (2003)
• [Publications] Nishiyama, N., et al.: "Differential gene expression profile between PC-14 cells treated with free cisplatin and cisplatin-incorporated polymeric micelles"Bioconjug.Chem.. 14・2. 449-457 (2003)
• [Publications] Tsunoda, T., et al.: "Differential gene expression profiles and identification of the genes relevent to clinicopathologic factors in colorectal cancer selected by cDNA array method in combination with principal component analysis"Int.J.Oncol.. (In press).
• [Publications] Kanzawa, F., et al.: "In vitro effects of combinations of cis-amminedichloro (2-methylpyridine) platinium (II) (ZD0473) with other movel anticancer drugs on the growth of SBC-3, a human small cell lung cancer cell line"Lung Cancer. (In press).
• [Publications] Koh, Y., et al.: "Mechanisms of action of cancer chemotherapeutic Agents : topoisomerase inhibitors.In "Cancer Handbook""Reference Nature Pulishing Group. 5 (2002)
• [Publications] Yoshida, M., et al.: "Antitumor action of the PKC activator gnidimacrin through cdk2 inhibition"Int. J. Cancer. 94・3. 348-352 (2001)
• [Publications] Zhang, J., et al.: "Differential osteopontin expression in lung cancer"Cancer Lett.. 171・2. 215-222 (2001)
• [Publications] Saijo, N., et al.: "New strategies for cancer therapy in the 21st century"Cancer Chemother. Pharmacol.. 48・Suppl 1. S102-106 (2001)
• [Publications] Yoshida, M., et al.: "Induction of MRP5 and SMRP mRNA by adriamycin exposure and its overexpression in human lung cancer cells resistant to adriamycin"Int. J. Cancer. 94・3. 432-437 (2001)
• [Publications] Fukuoka, K., et al.: "Mechanism of the radiosensitization induced by vinorelbine in human non-small cell lung cancer cells"Lung Cancer. 34・3. 451-460 (2001)
• [Publications] Tatsumi, Y., et al.: "Enhancement of in vivo antitumor activity of a novel antimitotic 1-phenylpropenone derivative, AM-132, by tumor necrosis factor-α or interleukin-6"Jpn. J. Cancer Res.. 92・7. 768-777 (2001)
• [Publications] Usuda, J., et al.: "Increased cytotoxic effects of photodynamic therapy in IL-6 gene transfected cells via enhanced apoptosis"Int. J. Cancer. 93・4. 475-480 (2001)
• [Publications] Natsume, T., et al.: "Gene expression profiling of exposure to TZT-1027, a novel microtubule-interfering agent in non-small cell lung cancer PC-14 cells and astrocytes"Invest. New Drugs. 19・4. 293-301 (2001)
• [Publications] Nakamura, T., et al.: "Reversal of cisplatin resistance by the 1,4-benzothiazepine derivative, JTV-519"Jpn. J. Cancer Res.. 92・6. 597-602 (2001)
• [Publications] Kanzawa, F., et al.: "In vitro synergistic interactions between the cisplatin analogue nedaplatin and the DNA topoisomerase I inhibitor irinotecan and the mechanism of this interaction"Clin. Cancer Res.. 7・1. 202-209 (2001)
• [Publications] Tsunoda, T., et al.: "Upregulated expression of angiogenesis genes and down regulation of cell cycle genes in human colorectal cancer tissue determined by cDNA macroarray"Anticancer Res.. 21・1A. 137-144 (2001)
• [Publications] Shimizu, M., et al.: "CPT-11 alters the circadian rhythm of dihydropyrimidine dehydrogenase mRNA in mouse liver"Jpn. J. Cancer Res.. 92・5. 554-561 (2001)
• [Publications] Suzuki, T., et al.: "The MRP family and anticancer drug metabolism current drug"Met. Curr. Drug Met.. 2・4. 367-377 (2001)
• [Publications] Fukuoka, K., et al.: "Mechanisms of action of the novel sulfonamide anticancer agent E7070 on cell cycle progression in human non-small cell lung cancer cells"Invest. New Drugs. 19・3. 219-227 (2001)
• [Publications] Nishio, K. et al.: "Oncoprotein 18 overexpression increases the sensitivity to vindesine in the human lung carcinoma cells"Cancer. 91・8. 1494-1499 (2001)
• [Publications] Natsume, T., et al.: "Enhanced antitumor activities of TZT-1027 against TNF-α or IL-6 secreting Lewis lung carcinoma in vivo"Cancer Chemother. Pharmacol.. 49・1. 35-47 (2002)
• [Publications] Koh, Y., et al.: "Decreased expression of α2,8 sialyltransferase and increased expression of β1,4 N-acetylgalactosaminyltransferase in gastrointestinal cancers"Proc. Soc. Exp. Biol. Med.. 227・3. 196-200 (2002)
• [Publications] Naruse, I., et al.: "Antitumor activity of the selective epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI)Iressa^<TM>(ZD1839) in a EGFR-expressing multidrug resistant cell line in vitro and in vivo"Int. J. Cancer. 98・2. 310-315 (2002)
• [Publications] Naruse, I., et al.: "Enhanced anti-tumor effect of trastuzumab in combination with cisplatin"Int. J. Cancer. (In press).
• [Publications] Fukuoka, K., et al.: "Mechanism of vinorelbine-induced radiosensitization of human small cell lung cancer cells"Cancer Chemother. Pharmacol.. (In press).
• [Publications] Takahashi, F., et al.: "Osteopontin induces angiogenesis of murine neuroblastoma cells in mice"Int. J. Cancer. (In press).
• [Publications] Kanzawa, F., et al.: "In vitro effects of combinations of cis-amminedichloro(2-methylpyridine)platinum (α)(ZD0473) with other novel anticancer drugs on the growth of SBC-3, a human small cell lung cancer cell line"Lung Cancer. (In press).
• [Publications] Koh, Y., et al.: "Mechanisms of action of cancer chemotherapeutic agents : topoisomerase inhibitors. In "Cancer Handbook""Reference Nature Publishing Group. 5 (2002)
• [Publications] Usuda,J., et al: "Molecular determinants of UCN-01 induced growth inhibition in human lung cancer cells."International Journal of Cancer. 85・2. 275-280 (2000)
• [Publications] Fukuoka,K., et al.: "Ectopic p16^<INK4> expression enhances CPT-11-induced apoptosis through the increased delay in S phase progression in human non-small cell lung cancer cells."International Journal of Cancer. 86・2. 197-203 (2000)
• [Publications] Fukumoto,H., et al.: "Activation-induced apoptosis of peripheral lymphocytes treated with 7-hydroxystaurosporine, UCN-01."Invest.New.Drugs. 17・4. 335-341 (2000)
• [Publications] Oguri,T. et al.: "Increased expression of the MRP5 gene is associated with exposure to platinum drugs in lung cancer."International Journal of Cancer. 86・1. 95-100 (2000)
• [Publications] Fukuoka,K., et al.: "Mechanism of action of aragusterol A (YTA0040), a potent antitumor marine steroid targeting the Gl phase of the cell cycle."International Journal of Cancer. 88・5. 810-819 (2000)
• [Publications] Kuh,H.-J., et al.: "A novel pharmacodynamic model for quantitative analysis of cell cycle arrest and its contribution to overall growth inhibition by anticancer agents."Jpn.J.Cancer Res.. 91・12. 1303-1313 (2000)