1995 Fiscal Year Annual Research Report

酵母人工染色体を用いた多剤耐性(MDR)遺伝子群の構造と機能に関する共同研究

Research Project

Project/Area Number	06044185
Research Institution	Kyushu University
Principal Investigator	河野公俊九州大学, 医学部, 助教授 (00153479)
Co-Investigator(Kenkyū-buntansha)	桑野信彦九州大学, 医学部, 教授 (80037431) 小野真弓九州大学, 医学部, 講師 (80128347) 和田守正九州大学, 医学部, 助手 (20220965) COLE Susan P カナダクィーンズ大学, 医学部, 教授 DEELEY Roger カナダクィーンズ大学, 癌研究所, 所長 FOJO Antonio 米国国立癌研究所, 主任 GREEN Eric D 米国国立衛生研究所, 主任
Keywords	酵母人工染色体 / 多剤耐性 / 第7染色体長腕21 / 遺伝子発現 / 遺伝子増幅 / 多剤耐性関連蛋白 / FISH / コンティングマップ
Research Abstract	多剤耐性遺伝子を含むヒト7番染色体長腕について、約30の人工酵母染色体(YAC)クローンをすでに分離していた。平成7年度は、そのゲノム構造解析の研究成果として、約1.5Mbに渡る領域のコンティングマップの作成を行い、成果としてまとめ国際誌に発表した。内容として、2つの多剤耐性遺伝子、MDR1とMDR3の染色体上の位置関係を明かにした。DNAプローブとSTSを用いて、多剤耐性細胞における、本領域の遺伝増幅の領域について、大まかに決定した。制限酵素による解析から、6個のCpG領域の存在を証明した。機能解析の研究効果としては、YACクローンのマウスL細胞へ導入し、薬剤耐性獲得における、導入遺伝しの発現の誘導に成功し、同じく国際誌に発表した。この発現により遺伝子産物は薬剤の細胞外への排出能をもつP糖蛋白が関与すること、それがマウスの形質膜に正しく発現したことをヒト特異的抗体を用いて証明した。又、その蛍光免疫染色の結果は、国際誌の表紙を飾ることとなった。以上の結果は、導入遺伝子が、特異的に、薬剤耐性獲得過程において、発現及び遺伝子増幅がしやすい状態を保持していることを示している。その原因の一つとして、マロマチン構造や、メチル化が関連すると考えられており、現在研究を続行中である。また、YACクローンが非常に大きな遺伝子の機能解析に最適であることから、LCRの存在などを明かにする上でこの導入実験はよいモデル系となることも示している。

Research Products

(16 results)

All Other

All Publications (16 results)

[Publications] K.Kohno: "A novel mutation of DNA topoisomerase II α gene in an etoposide-resistant humen cancer cell line." Cell.Pharmacol.2. 87-90 (1995)
[Publications] K.Samoto: "Expression of vascular endothelial growth factor and its possible relation with neovascularization in human brain tumors." Cancer Res.55. 1189-1193 (1995)
[Publications] T.Abe: "Chemosensitization of spontaneous multidrug resistance by a 1,4-dihydropyridine analog and verapamil in human glioma cell lines overexpressing MRP or MDR1." Brit.J.Cancer. 72. 418-423 (1995)
[Publications] S.Sato: "Differential effects of protein kinase inhibitor,H-7,on activation of human multidrug resistance 1 (MDR1) promoter." Cell.Pharmacol.2. 153-157 (1995)
[Publications] S.Jimi: "Modulation by bovine angiogenin of tubular morphogenesis and expression of plasminogen activator in bovine endothelial cells." Biochem.Biophys.Res.Commun.211. 476-483 (1995)
[Publications] A.Yokomizo: "Markedly decerased expression of glutathione S-transferase-π gene in human cancer cell lines resistant to buthionine sulfoximine,an inhibitor of cellular glutathione synthesis." J.Biol.Chem.270. 19451-19457 (1995)
[Publications] T.Kubo: "DNA topoisomerase II α gene expression under transcriptional control in etoposide/teniposide resistant human cancer cells." Cancer Res.55. 3860-3864 (1995)
[Publications] A.Yokomizo: "Cellular levels of thioredoxin associated with drug sensitivity to cisplatin,mitomycin C,doxorubicin and etoposide." Cancer Res.55. 4293-4296 (1995)
[Publications] Y.Hirohashi: "Bimodulation by hyperthermia of topoisomerase II-targeting drugs in human clorectal cancer cells." Jpn.J.Cancer Res.86. 1097-1105 (1995)
[Publications] K.Torigoe: "YAC-based 1.5Mb contig spanning the human multidrug resistantce gene region and delineating the amplification unit in two human multidrug resistant cell lines." Genome Research. 5. 233-244 (1995)
[Publications] H.Kusaba: "Functional expression of yeast artificial chromosome (YAC) human multidrug resistance genes in mouse cells." Genome Research. 5. 245-258 (1995)
[Publications] K.Asakuno: "The exogenous control of transfected c-fos gene expression and angiogenesis in cells implanted inot the rat brain." Brain.Res.702. 23-31 (1995)
[Publications] S.Ushiro: "Heparin-binding EGF-like growth factor : p91 activation,induction of plasminogen activator/plasminogen activator inhibitor. and tubular morphogenesis in human microvascular endothelial cells." Jpn.J.Cancer Res.87. 68-77 (1996)
[Publications] T.Okimoto: "Regulation of epidermal growth factor receptor by activated H-ras and v-myc oncogenes in mouse balb/3T3 cells : possible roles of AP-1." Oncogene. 12 (in press). (1996)
[Publications] T.Shono: "Involvement of a transcription factor NFkB in tubular morphogenesis of human microvascular endothelial cells by oxidative stress." Molec.Cell Biol.(in press). (1996)
[Publications] M.Ono: "Angiogenesis as a new target for cancer treatment." Cancer Chemotherapy Challenges for the Future (eds.By K.Kimura,K.Ota,K.Yamada,H.Saito and M.Ogawa) Vol.10,Excerpta Medica. (in press). (1996)

1995 Fiscal Year Annual Research Report

酵母人工染色体を用いた多剤耐性(MDR)遺伝子群の構造と機能に関する共同研究

Principal Investigator

河野 公俊 九州大学, 医学部, 助教授 (00153479)

Research Products

[Publications] K.Kohno: "A novel mutation of DNA topoisomerase II α gene in an etoposide-resistant humen cancer cell line." Cell.Pharmacol.2. 87-90 (1995)

[Publications] K.Samoto: "Expression of vascular endothelial growth factor and its possible relation with neovascularization in human brain tumors." Cancer Res.55. 1189-1193 (1995)

[Publications] T.Abe: "Chemosensitization of spontaneous multidrug resistance by a 1,4-dihydropyridine analog and verapamil in human glioma cell lines overexpressing MRP or MDR1." Brit.J.Cancer. 72. 418-423 (1995)

[Publications] S.Sato: "Differential effects of protein kinase inhibitor,H-7,on activation of human multidrug resistance 1 (MDR1) promoter." Cell.Pharmacol.2. 153-157 (1995)

[Publications] S.Jimi: "Modulation by bovine angiogenin of tubular morphogenesis and expression of plasminogen activator in bovine endothelial cells." Biochem.Biophys.Res.Commun.211. 476-483 (1995)

[Publications] A.Yokomizo: "Markedly decerased expression of glutathione S-transferase-π gene in human cancer cell lines resistant to buthionine sulfoximine,an inhibitor of cellular glutathione synthesis." J.Biol.Chem.270. 19451-19457 (1995)

[Publications] T.Kubo: "DNA topoisomerase II α gene expression under transcriptional control in etoposide/teniposide resistant human cancer cells." Cancer Res.55. 3860-3864 (1995)

[Publications] A.Yokomizo: "Cellular levels of thioredoxin associated with drug sensitivity to cisplatin,mitomycin C,doxorubicin and etoposide." Cancer Res.55. 4293-4296 (1995)

[Publications] Y.Hirohashi: "Bimodulation by hyperthermia of topoisomerase II-targeting drugs in human clorectal cancer cells." Jpn.J.Cancer Res.86. 1097-1105 (1995)

[Publications] K.Torigoe: "YAC-based 1.5Mb contig spanning the human multidrug resistantce gene region and delineating the amplification unit in two human multidrug resistant cell lines." Genome Research. 5. 233-244 (1995)

[Publications] H.Kusaba: "Functional expression of yeast artificial chromosome (YAC) human multidrug resistance genes in mouse cells." Genome Research. 5. 245-258 (1995)

[Publications] K.Asakuno: "The exogenous control of transfected c-fos gene expression and angiogenesis in cells implanted inot the rat brain." Brain.Res.702. 23-31 (1995)

[Publications] S.Ushiro: "Heparin-binding EGF-like growth factor : p91 activation,induction of plasminogen activator/plasminogen activator inhibitor. and tubular morphogenesis in human microvascular endothelial cells." Jpn.J.Cancer Res.87. 68-77 (1996)

[Publications] T.Okimoto: "Regulation of epidermal growth factor receptor by activated H-ras and v-myc oncogenes in mouse balb/3T3 cells : possible roles of AP-1." Oncogene. 12 (in press). (1996)

[Publications] T.Shono: "Involvement of a transcription factor NFkB in tubular morphogenesis of human microvascular endothelial cells by oxidative stress." Molec.Cell Biol.(in press). (1996)

[Publications] M.Ono: "Angiogenesis as a new target for cancer treatment." Cancer Chemotherapy Challenges for the Future (eds.By K.Kimura,K.Ota,K.Yamada,H.Saito and M.Ogawa) Vol.10,Excerpta Medica. (in press). (1996)

河野公俊九州大学, 医学部, 助教授 (00153479)