Development of real-time intracellular pH sensor using a membrane-permeable peptide nucleic acid.

2005 Fiscal Year Final Research Report Summary

• Project/Area Number: 16500200
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Neuroscience in general
• Research Institution: Okayama University
• Principal Investigator: TOMIZAWA Kazuhito Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Associate Prof., 大学院・医歯薬学総合研究科, 助教授 (40274287)
• Co-Investigator(Kenkyū-buntansha): MATSUSHITA Masayuki Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Assistant Prof., 大学院・医歯薬学総合研究科, 講師 (30273965) ; MATSUI Hideki Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (30157234)
• Project Period (FY): 2004 – 2005
• Keywords: Peptide nucleic acid / Polyarginine / Nucleic acid / Bio-molecule / Bio-imaging / Cell

Research Abstract

Membrane-permeable peptide nucleic acid (PNA) developed by us is thought to be a powerful tool for the regulation of cell function. The purpose of this project is that development of the tools that are useful for the regulation of protein expression and the function and for the monitoring of nano-ecology in living cells using the PNA. In the present project, we show that membrane-permeable PNA is a powerful tool for the regulation of gene expression and for anti-cancer therapy as follows.
1.Regulation of gene expression.
We synthesized a PNA, which effectively hybridized with plasmid DNA, and the PNA was fused a membrane-permeable poly-arginine to have the ability of membrane permeability. After mixing the PNA with plasmid DNAs, cells were incubated with the complex and the efficiency of the transfection was investigated. The plasmid DNAs were effectively transfected in both cells and the efficiency is better than that of cationic transfection method.
2.Antigene therapy using membrane-permeable PNA.
We synthesized PNAs that are able to bind with p53 mRNA and the double-strand DNA. Cancer cells were incubated with the PNAs. The PNAs were delivered into cells and inhibited the expression of p53 mRNA and the protein. The combination treatment of the PNAs with p53 protein therapy was capable to replace endogenous mutated p53 proteins with wild-type p53 proteins in the cancer cells.

Research Products

• [Journal Article] p53 protein transduction therapy : successful targeting and inhibition of the growth of the bladder cancer cells. (2006)
  • Author(s): Inoue, M., Tomizawa, K.et al.
  • Journal Title: Eur. Urol. 49 ・ 1 Pages: 161-168
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] p53 protein transduction therapy : Successful targeting and inhibition of the growth of the bladder cancer cells. (2006)
  • Author(s): Inoue, M., Tomizawa, K., Matsushita, M., Lu, Y.-F., Yokoyama, T., Yanai, H., Takashima, A., Kumon, H., Matsui, H.
  • Journal Title: Eur.Urol. 49・1 Pages: 161-168
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The NH-2-terminal of influenza virus hemagglutinin-2 subunit peptides enhance the anti-tumor potency of poly-arginine-mediated p53 protein transduction. (2005)
  • Author(s): Michiue, H., Tomizawa, K.et al.
  • Journal Title: J. Bio1. Chem. 280 ・ 9 Pages: 8285-8289
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Ubiquitination-resistant p53 protein transduction therapy facilitates anti-cancer effect on the growth of human malignant glioma cells. (2005)
  • Author(s): Michiue, H., Tomizawa, K.et al.
  • Journal Title: FEBS Lett. 579 ・ 18 Pages: 3965-3969
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Cdk5-dependent regulation of glucose-stimulated insulin secretion. (2005)
  • Author(s): Wei, F.-Y., Tomizawa, K.et al.
  • Journal Title: Nature Med. 11 ・ 10 Pages: 1104-1108
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Control of cyclin-dependent kinase 5 (Cdk5) activity by glutamatergic regulation of p35 stability. (2005)
  • Author(s): Wei, F.-Y., Tomizawa, K.et al.
  • Journal Title: J. Neurochem. 93 ・ 2 Pages: 502-512
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Truncation and activation of calcineurin A by calpain I in Alzheimer disease brain. (2005)
  • Author(s): Liu, F., Tomizawa, K.et a1.
  • Journal Title: J. Bio1. Chem. 280 ・ 45 Pages: 37755-37762
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The NH-2-terminal of influenza virus hemagglutinin-2 subunit peptides enhance the anti-tumor potency of poly-arginine-mediated p53 protein transduction. (2005)
  • Author(s): Michiue, H., Tomizawa, K., Wei, F.-Y., Matsushita, M., Lu, Y.-F., Ichikawa, T., Tamiya, T., Date, I., Matsui, H.
  • Journal Title: J.Biol.Chem. 280・9 Pages: 8285-8289
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Ubiquitination-resistant p53 protein transduction therapy facilitates anti-cancer effect on the growth of human malignant glioma cells. (2005)
  • Author(s): Michiue, H., Tomizawa, K., Matsushita, M., Tamiya, T., Lu, Y.-F., Ichikawa, T., Date, I., Matsui, H.
  • Journal Title: FEBS Lett. 579・18 Pages: 3965-3969
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cdk5-dependent regulation of glucose-stimulated insulin secretion. (2005)
  • Author(s): Wei, F.-Y., Nagashima, K., Ohshima, T., Saheki, Y., Lu, Y.-F., Matsushita, M., Yamada, Y., Mikoshiba, K., Seino, Y., Matsui, H., Tomizawa, K.
  • Journal Title: Nature Med. 11・10 Pages: 1104-1108
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Control of Cyclin-dependent kinase 5 (Cdk5) activity by glutamatergic regulation of p35 stability. (2005)
  • Author(s): Wei, F.-Y., Tomizawa, K., Ohshima, T., Asada, A., Saito, T., Nguyen C., Bibb, JA., Ishiguro, K., Kulkami, A.B., Pant, H.C., Mikoshiba, K., Matsui, H., Hisanaga, S.
  • Journal Title: J.Neurochem. 93・2 Pages: 502-512
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Truncation and activation of calcineurin A by calpain I in Alzheimer disease brain. (2005)
  • Author(s): Liu, F., Grundke-Iqbal, I., Iqbal, K., Oda, Y., Tomizawa, K., Gong C.-X.
  • Journal Title: J.Biol.Chem. 280・45 Pages: 37755-37762
  • Description: 「研究成果報告書概要(欧文)」より