Organotypic epithelial raft cultures as HPV-related cancer models for evaluating siRNA and its delivery system

2012 Fiscal Year Final Research Report

• Project/Area Number: 22592084
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pathobiological dentistry/Dental radiology
• Research Institution: University of Tsukuba (2012); Tokyo Medical and Dental University (2010-2011)
• Principal Investigator: YAMATO Kenji 筑波大学, 医学医療系, 研究員 (50174751)
• Project Period (FY): 2010 – 2012
• Keywords: HPV / RNAi / ラフトカルチャー

Research Abstract

To utilize siRNA technology to the treatment of HPV-related diseases, developing an efficient siRNA delivery system is essential. In this study, we established organotypic raft cultures using TERT- and HPV16-E6E7 immortalized human keratinocytes stably expressing firefly luciferase. These cultures facilitated quantitative analysis of knock-down efficiency induced by local siRNA delivery and showed that some cationic liposomes could deliver siRNA through stratified squamous epithelium.

Research Products

• [Journal Article] The E1 protein of human papillomavirus type 16 is dispensable for maintenance replication of the viral genome (2012)
  • Author(s): Egawa, N., Nakahara, T., Ohnoa,S., Narisawa-Saitoa,M., Yugawa,T., Fujita,M. Yamato, K., Natori Y. and Kiyono, T
  • Journal Title: J Virol Volume: 86 Pages: 3276-3283
  • DOI: DOI:10.1128/JVI.06450-11
  • Peer Reviewed
• [Journal Article] Enhanced specificity of HPV16 E6E7 siRNA by RNA-DNA chimera modification (2011)
  • Author(s): Yamato, K., Egawa, N., Endo, S., Ui-Tei, K., Yamada, T., Saigo, K., Hyodo, I., Kiyono, T. and Nakagawa, I
  • Journal Title: Cancer Gene Ther Volume: 18 Pages: 587-597
  • DOI: DOI:10.1038/cgt.2011.28
  • Peer Reviewed
• [Journal Article] Potent in vitro and in vivo antitumor effects of MDM2 inhibitor nutlin-3 in gastric cancer cells (2011)
  • Author(s): Endo, S., Yamato, K., Hirai, S., Moriwaki, M., Fukuda, K., Suzuki, H., Abei, M., Nakagawa, I. and Hyodo, I
  • Journal Title: Cancer Sci Volume: 102 Pages: 605-613
  • DOI: DOI:10.1111/j.1349-7006.2010.01821
  • Peer Reviewed
• [Presentation] Identification of a short RNA segment in the siRNA seed region required for efficient RNAi (2012)
  • Author(s): 大和建嗣
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡国際会議場・マリンメッセ福岡 (福岡市)
  • Year and Date: 20121211-14
• [Presentation] ヒト胃癌細胞に対するclassIII HDAC阻害剤 tenovin-6の抗腫瘍効果に関する検討 (2012)
  • Author(s): 廣瀬充明
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: ロイトイン札幌(札幌
  • Year and Date: 20120919-21
• [Presentation] The antitumor effect of HDAC inhibitor Tenovin-6 for gastric cancer cells (2012)
  • Author(s): Ueno T
  • Organizer: American Association of Cancer Ressearch 103rd Annual Meeting
  • Place of Presentation: Chicago, IL
  • Year and Date: 20120331-0401
• [Presentation] AGO2-association, microRNA suppression and cytotoxicity ofRNA-DNA chimera modified siRNA (2011)
  • Author(s): 大和建嗣
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(横浜市)
  • Year and Date: 20111213-16
• [Presentation] The anti-tumor effect of MDM2 antagonist nutlin-3 in gastric cancer cell in vitro and in vivo (2010)
  • Author(s): Endo S
  • Organizer: American Association of Cancer Ressearch 101st Annual Meeting
  • Place of Presentation: Washington, DC
  • Year and Date: 20100417-21