Search and analysis of genes involved in the sensitivity to arsenic trioxide of human cells.

• Project/Area Number: 15590277
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pathological medical chemistry
• Research Institution: Kagoshima University
• Principal Investigator: AKIYAMA Shin-ichi Kagoshima University, Graduate School of Medial and Dental Sciences, Professor, 大学院・医歯学総合研究科, 教授 (60117413)
• Co-Investigator(Kenkyū-buntansha): FURUKAWA Tatsuhiko Kagoshima University, Graduate School of Medial and Dental Sciences, Associate Professor, 大学院・医歯学総合研究科, 助教授 (40219100)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000); Fiscal Year 2003: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: ABCB6 / resistance / cisplatin / arsenite / mitochondria / MRP1 / signature sequence / GSH / ヒ素 / 耐性 / シスプラチン / ABCトランスポーター / ABCトランスポータ

Research Abstract

1.We have isolated aesenite-resistant human epidermoid carcinoma KB cells, termed KAS. KAS cells are resistant to both sodium arsenite and cisplatin compared to the parental KB-3-1 cells. To identify the genes responsible for the resistance to these agents, we employed microarray analysis, using a platform dedicated to drug resistance. The ABC-ToxChip and real-time PCR analyses revealed the increased expression of the mitocondrial ABC transporter ABCB6 in KAS cells. We established KB-B6 cells, KB-3-1 cells stably transfected with ABCB6 expression vector, to study the role of the transporter in arsenite resistance. KB-B6 cells were resistant to sodium arsenite and Accurnulation of cisplatin in KB-B6 cisplatin in KB-B6 cells was decreassed compared with the KB/CV cells cisplatin.
The decleased DNA adduct in both genome and mitochondrial DNAs in KB-B6 cells was observed. Our results suggest that ABCB6 is also localized in plasma membranes and transports cisplatin outside the cells.
2.We studied the mechanism of drug transport by MRP1 that confers resistance to arsenite. We investigated the function of the signature sequences in NBDs of MRP1 using the mutant MRP1s that have mutations of the signature sequences (G771D and G1433D). Our findings suggested that both signature sequences of MRP1 are involved in ATP hydrolysis and must be intact for the ATP hydrolysis and transport by MRP1.
The accumulated evidence has proved that GSH interacts with MRP1 and stimulates drug transport. However, the mechanism of GSH-dependent drug trasport by MRP1 remains unclear. In this study, we used limited tryptic digestion of MRP1 in isolated membrane vesicles, in the presence and absence of GSH, to investigate the influence of GSH on MRP1 conformation. Our results suggested that GSH induces a conformational change at a site within the MRP1 that is indispensable for the interaction of MRP1 with its substrates.

Research Products

• [Journal Article] GSH suppresses a tryptic site in the C-terminal half of human MRP1. (2005)
  • Author(s): Ren, X.-Q.
  • Journal Title: J.Biol.Chem. 280 Pages: 6231-6237
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Cepharanthine potently enhances the sensitivity of anticancer Agents in K562 cells (2005)
  • Author(s): Ikeda, R., Che, X-F, Yamaguchi, T., Ushiyama, M., Zeng, C-L., Okumura, H., Takeda, Y, Shibayama, Y, Nakamura, K., Jeung, H-C., Furukawa, T., Sumizawa, T., Haraguchi, M., Akiyama.S., Yamada K.
  • Journal Title: Cancer Sci 96 Pages: 327-376
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] GSH suppresses a tryptic site in the C-terminal half of human MRP1. (2005)
  • Author(s): Ren, X-Q., Furukawa, T., Nakajima, Y, Takahasi, Aoki, H., Sumizawa, T., Haraguchi, M., Kobayashi, M., Chijiwa, K., Akiyama, S.
  • Journal Title: J.Biol.Chem. 280 Pages: 6231-6237
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Inhibition of metastasis of tumor cells overexpressing thymidine (2004)
  • Author(s): Nakajima, Y.
  • Journal Title: Cancer Res. 64 Pages: 1749-1801
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Function of the ABC signature sequences in the human multidrug resistance protein 1. (2004)
  • Author(s): Ren, X.-Q.
  • Journal Title: Mol.Pharmacol 65 Pages: 1536-1542
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Kendarimide A, a novel peptide reversing P-glycoprotein-mediated multidrug resistance in tumor cells, from a marine sponge of Haliclona SP. (2004)
  • Author(s): Aoki, S.
  • Journal Title: Tetrahedron 60 Pages: 7053-7059
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Induction of cellular resistance to nucleoside reverse transcriptase inhibitors by the wild-type breast cancer resistance protein. (2004)
  • Author(s): Wang, X.
  • Journal Title: Biochem Pharmacol. 68 Pages: 1363-1370
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Characterization of MVP and VPARP assembly into vault ribonucleoprotein complexes. (2004)
  • Author(s): Zheng, C.-L.
  • Journal Title: Biochem.Biophys.Res.Commun. 326 Pages: 100-107
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Characterization of MVP and VPARP assembly into vault ribonucleoprotein complexes. (2004)
  • Author(s): Zheng, C-L., Sumizawa, T., Che, X-F, Tsuyama, S., Furukawa, T., Haraguchi, M., Gao, H., Gotanda, T., Jueng, H-Cl., Murata, F., Akiyama, S.
  • Journal Title: Biochem.Biophys.Res.Commun. 326 Pages: 100-107
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Function of the ABC signature sequences in the human multidrug resistance protein 1. (2004)
  • Author(s): Ren, X.-Q., Furukawa, T., Haraguchi, M., Sumizawa, T., Aoki, S., Kobayashi, M., Akiyama, S.
  • Journal Title: Mol.Pharmacol. 65 Pages: 1536-1542
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Inhibition of Metastasis of tumor cells overexpressing theymidine phosphorylase by 2-Deoxy-L-ribose. (2004)
  • Author(s): Nakajima, Y., Gotanda, T., Uchimiya, H., Furukawa, T., Haraguchi, M., Ikeda, R., Sumizawa, T., Yoshida, H., Akiyama, S.
  • Journal Title: Cancer Res. 64 Pages: 1749-1801
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Breast cancer resistance protein (BCRP/ABCG2) induces cellular resistance to HIV-1 nucleoside reverse transcriptase inhibitors. (2003)
  • Author(s): Wang, X., Furukawa, T., Nitanda, T., Okamoto, M., Sugimoto, Y., Akiyama, S., Baba, M.
  • Journal Title: Mol.Pharmacol. 63 Pages: 65-72
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Localization of the GSH-dependent photolabelling site of an agosterol A analog on human MRP1. (2003)
  • Author(s): Ren, X.-Q., Furukawa, T., Aoki, S., Sumizawa, T., Haraguchi, M., Che, X-F., Kobayashi, M., Akiyama, S.
  • Journal Title: Br.J.Pharmacol. 138 Pages: 1553-1561
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Thymidine phosphorylase inhibits apoptosis induced by cisplatin. (2003)
  • Author(s): Ikeda, R., Furukawa, T., Mitsuo, R., Noguchi, T., Kitazono, M., Okumura, H., Sumizawa, T., Haraguchi, M., Che, X-F, Uchimiya, H., Nakajima, Y., Ren, X-Q., Oiso, S., Inoue, I., Yamada, K., Akiyama, S.
  • Journal Title: Biochem.Biophys.Res.Commun. 301 Pages: 358-363
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Thymidine phosphorylase suppresses apoptosis induced by Microtubule-interfering agents.
  • Author(s): Jeung, H-C., Che, X-F, Haraguchi, M., Furukawa, T., Zeng, C-L., Sumizawa, T., Rha, S-Y, Akiyama, S.
  • Journal Title: Biochemical Pharmacol. (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Nakajima Y: "Inhibition of metastasis of tumor cells overexpressing phosphorylase by 2-deoxy-L-ribose"Cancer Res.. (In Press).
• [Publications] Wang X: "Breast cancer resistance protein(BCRP/ABCG2) induces cellular reistance to HIV-1 nucleoside reverse transcriptase inhibitors"Mol.Pharmacol.. 63. 65-72 (2003)
• [Publications] Mukai M: "Reversal of the resistance to STI571 in human chronic myelogenous leukemia K562 cells."Cancer Sci.. 94. 557-563 (2003)
• [Publications] Ren X-Q: "Localization of the GSH-dependent photolabelling site of an agosterol A analog on human MRP1."Br.J.Pharmacol.. 138. 1553-1561 (2003)
• [Publications] Ikeda R: "Thymidine phosphorylase inhibits apoptosis induced by cisplatin."Biochem Biophys Res Commun.. 301. 358-363 (2003)
• [Publications] Mitsuo M: "Binding site(s) on P-glycoprotein for a newly synthesized photoaffinity analog of agosterol A."Oncol Res.. 14. 39-48 (2003)
• [Publications] 秋山伸一 (共著): "臨床腫瘍学"癌と化学療法社. 2031 (2003)