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

• 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
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2011: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2010: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: HPV / RNAi / ラフトカルチャー / RNA干渉 / siRNA / 子宮頸癌 / GFP / 皮膚 / ルシフェラーゼ

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., et al
  • Journal Title: J. Virol. Volume: 86 Pages: 3276-3281
  • DOI: 10.1128/jvi.06450-11
  • Peer Reviewed
• [Journal Article] Enhanced specificity of HPV16 E6E7 siRNA --- (2011)
  • Author(s): Yamato K. et al.
  • Journal Title: Cancer Gene Therapy Volume: 18 Pages: 587-597
  • 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
  • NAID: 10029291334
  • 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.
  • Journal Title: Cancer Sci. Volume: 102 Pages: 605-613
  • 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)
• [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
• [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
• [Presentation] Identification of a short RNA segment in the siRNA seed region required for efficient RNAi (2012)
  • Author(s): 大和建嗣（代表）、遠藤慎治
  • 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）
• [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（代表）、遠藤慎治、齋藤梨絵、上野 卓教、鈴木英雄、大和 建嗣、兵頭一之介
  • 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）
• [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)
• [Presentation] Enhanced specificity of HPV16 E6E7 siRNA by RNA-DNA chimera modification (2011)
  • Author(s): Yamato, K
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: 横浜
• [Presentation] Enhanced specificity of HPV16 E6E7 siRNA by RNA-DNA chimera modification. (2010)
  • Author(s): Yamato, K.
  • Organizer: 第33回日本分子生物学会年会
  • Place of Presentation: 神戸
  • Year and Date: 2010-12-08
• [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
• [Presentation] The anti-tumorcancer 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 Research 101st Annual Meeting 2010
  • Place of Presentation: Washington, DC