Establishment of oral cancer stem-like cells using normal human tongue keratinocytes

2013 Fiscal Year Final Research Report

• Project/Area Number: 24792264
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Surgical dentistry
• Research Institution: Hyogo Medical University
• Principal Investigator: ZUSHI Yusuke 兵庫医科大学, 医学部, 助教 (80581206)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Keywords: ヒト正常舌角化細胞 / 口腔がん幹細胞

Research Abstract

We newly established human tongue keratinocytes (HTK) cell lines derived from tongue mucosa with retroviral vectors expressing mutant CDK4, cyclinD1, hTERT and dominant negative form of p53. Additional transduction of oncogenic Hras or EGFR together with c-Myc into the HTK-K4DT-DNp53 showed subcutaneous tumor formation in nude mice only around 2 weeks. To determine the frequency of oral cancer initiating cells within these cells, a limiting dilution assay was performed. Orthotopic transplantation of HTK-K4DT-DNp53-Hras-cMyc resulted inlocal growth of tumors in all the mice tested and some of them yielded regional metstastases in 2-3 weeks.We have established an in vitro model for OSCCs with normal human tongue keratinocytes (HTKs) focusing on sequential transduction of defined genetic elements and succeeded in the creation of highly potent cancer initiating cells by introduction of c-MYC and oncogenic Hras on a background of the blockade of the pRB and p53 pathways .

Research Products

• [Journal Article] 口腔扁平上皮癌の発生における Human papillomavirus(HPV) 感染の関与と診断法の検討 (2013)
  • Author(s): 野口一馬, 寺田友紀, 中野芳郎, 山村倫世, 吉川恭平, 頭司雄介, 森寺邦康, 高岡一樹, 浦出雅裕, 岸本裕充
  • Journal Title: 日本口腔感染症学会雑誌 Volume: vol.20 No.2 Pages: 70-76
  • Peer Reviewed
• [Journal Article] Effect of a nitric oxide synthase inhibitor and a CXC chemokine receptor-4 antagonist on tumor growth and metastasis in a xenotransplanted mouse model of adenoid cystic carcinoma of the oral floor (2013)
  • Author(s): Takaoka K, Zushi Y, Noguchi K, Urade M, et al
  • Journal Title: Int J Oncol Volume: 43(3) Pages: 737-45
  • DOI: 10.3892/ijo.2013.2011
  • Peer Reviewed
• [Journal Article] Functional analysis of Zyxin in cell migration and invasive potential of oral squamous cell carcinoma cells (2013)
  • Author(s): Yamamura M, Noguchi K, Zushi Y, Urade M, et al
  • Journal Title: Int J Oncol Volume: 42(3) Pages: 873-80
  • DOI: 10.3892/ijo.2013.1761
  • Peer Reviewed
• [Journal Article] HPV 陽性および陰性の口腔扁平上皮がんに対する in vitro 多段階発がんモデル (2013)
  • Author(s): 頭司雄介, 野口一馬, 浦出雅裕
  • Journal Title: 日本口腔外科学会雑誌 Volume: vol.59 No.3 Pages: 13-25
  • Peer Reviewed
• [Presentation] in vitro 発癌モデルを用いた口腔発癌におけるドライバー変異の解明 (2013)
  • Author(s): 野口一馬, 頭司雄介, 吉川恭平, 山村倫世, 吉岡済, 浦出雅裕, 岸本裕充
  • Organizer: 日本口腔組織培養学会設立 50 周年記念学術大会
  • Place of Presentation: 東京
  • Year and Date: 2013-11-23
• [Presentation] 組織培養を用いた口腔癌研究の最前線-発がんから医療への応用- (2013)
  • Author(s): 野口一馬, 頭司雄介, 吉川恭平, 山村倫世, 吉岡済, 浦出雅裕, 岸本裕充
  • Organizer: 日本口腔組織培養学会設立50周年記念学術大会
  • Place of Presentation: 東京
  • Year and Date: 2013-11-23
• [Presentation] Establishment of oral cancer stem-like cells using normal human tongue keratinocytes (2013)
  • Author(s): Zushi Y, Noguchi K, Yamamura M, Moridera K, Takaoka K, Okui S, Urade M, Kishimoto H
  • Organizer: 54th Congress of the Korean Association of Oral Maxillofacial Surgery (KAOMS2013)
  • Place of Presentation: Daegu, South Korea
  • Year and Date: 2013-04-26
• [Presentation] 角化囊胞性歯原性腫瘍細胞の細胞株の樹立と病態モデル作成の試み (2012)
  • Author(s): 野口一馬, 藤井碧, 頭司雄介, 山村倫世, 吉川恭平, 岸本裕充, 浦出雅裕
  • Organizer: 第49回日本口腔組織培養学会学術大会
  • Place of Presentation: 広島
  • Year and Date: 2012-11-24
• [Presentation] 当科における口腔扁平上皮癌の臨床的検討- RADPLAT 導入前後の比較- (2012)
  • Author(s): 野口一馬, 吉川恭平, 瀬川英美, 川邊睦記, 頭司雄介, 山村倫世, 奥井森, 森寺邦康, 高岡一樹, 岸本裕充, 浦出雅裕
  • Organizer: 第57回日本口腔外科学会総会・学術大会
  • Place of Presentation: 横浜
  • Year and Date: 2012-10-19
• [Presentation] 病態モデル作成を目指した角化囊胞性歯原性腫瘍細胞の不死化の試みと細胞株の樹立 (2012)
  • Author(s): 野口一馬, 藤井碧, 頭司雄介, 山村倫世, 吉川恭平, 岸本裕充, 浦出雅裕
  • Organizer: 第57回日本口腔外科学会総会・学術大会
  • Place of Presentation: 横浜
  • Year and Date: 2012-10-19
• [Presentation] Analysis of carcinogenesis for both HPV-positive and - negative oral squamous cell carcinoma using normal human tongue keratinocytes (2012)
  • Author(s): Zushi Y, Noguchi K, Yoshikawa K, Okui S, Takaoka K, Moridera K, Kishimoto H, Urade M
  • Organizer: 21th Congress of the European Association for Cranio-MaxilloFacial Surgery (EACMFS 2012)
  • Place of Presentation: Dubrovnik, Croatia
  • Year and Date: 2012-09-12
• [Presentation] 口腔発がんにおける p53, EGFR, Ras 遺伝子発現の関連性について~ 不死化ヒト正常舌ケラチノサイト(HTK)を用いた解析~ (2012)
  • Author(s): 頭司雄介, 野口一馬, 吉川恭平, 岸本裕充, 浦出雅裕
  • Organizer: 第66回日本口腔科学会総会
  • Place of Presentation: 広島
  • Year and Date: 2012-05-18