Study on malignant transformation through DNA methylation in HPV16-positive oral squamous cell carcinoma

• Project/Area Number: 18K09791
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 57060:Surgical dentistry-related
• Research Institution: Hiroshima University
• Principal Investigator: SUGIYAMA Masaru 広島大学, 医系科学研究科(歯), 名誉教授 (70187681)
• Co-Investigator(Kenkyū-buntansha): 重石 英生 広島大学, 医系科学研究科(歯), 講師 (90397943)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: HPV16 / 癌幹細胞 / 口腔癌 / 歯周病 / 歯周病原細菌 / 口腔扁平上皮癌 / HPV / CD44 / メチル化

Outline of Final Research Achievements

①In HPV16 E6/E7-positive oral squamous cell carcinoma cell line (UM-SCC-104), E6/E7 siRNA knockdown significantly reduced sphere colony formation ability and mRNA expression of stem cell markers (Oct4, Nanog, ALDH1). ② Methylation of CD44 gene was detected more frequently in HPV16-positive oral cancer than in HPV16-negative oral cancer. ③ Characteristic oral bacterial flora was found in the subjects with HPV16 DNA. Additionally, there was a significant relationship between the presence of HPV16 DNA and bleeding during periodontal probing.

Academic Significance and Societal Importance of the Research Achievements

①HPV16陽性口腔扁平上皮癌細胞の幹細胞性の維持には、E6/E7遺伝子が重要な役割を持つことが示唆された。また、HPV DNAが宿主ゲノムに結合している子宮頸癌細胞とは異なり、UM-SCC-104細胞中ではHPV16 DNAがエピゾームとして存在しているにも関わらず、E6/E7 siRNA ノックダウンにより幹細胞性が低下した。このことは、存在様式に関わらず、HPV DNAが細胞の悪性化に関与していることを示唆している。②HPV16 DNA がCD44のメチル化に関与している可能性が示唆された。③歯周病とHPV感染との関係が示唆された。

Research Products

• [Journal Article] Detection of HPV16 E6 DNA in periodontal pockets of middle-aged and older people. (2021)
  • Author(s): Shigeishi H, Murodumi H, Ohta K, Sugiyama M.
  • Journal Title: Oral Science International. Volume: 18 Pages: 50-55
  • Peer Reviewed
• [Journal Article] Association of oral HPV16 infection with periodontal inflammation and the oral microbiome in older Japanese women. (2021)
  • Author(s): Shigeishi H, Su CY, Kaneyasu Y, Matsumura M, Nakamura M, Ishikawa M, Saito A, Ohta K, Sugiyama M.
  • Journal Title: Exp Ther Med. Volume: 21 Issue: 2 Pages: 167-167
  • DOI: 10.3892/etm.2020.9598
  • Peer Reviewed
• [Journal Article] Relationship between the prevalence of oral human papillomavirus DNA and periodontal disease (Review). (2021)
  • Author(s): Shigeishi H, Ohta K, Sugiyama M.
  • Journal Title: Biomedical Reports Volume: 14 Pages: 40-40
  • Peer Reviewed / Open Access
• [Journal Article] Detection of HPV16 E6 DNA in periodontal pockets of middle-aged and older people. (2021)
  • Author(s): Shigeishi H, Murodumi H, Ohta K, Sugiyama M.
  • Journal Title: Oral Sci Int Volume: 18 Issue: 1 Pages: 50-55
  • DOI: 10.1002/osi2.1079
  • Peer Reviewed
• [Journal Article] Relationship between the prevalence of oral human papillomavirus DNA and periodontal disease. (2021)
  • Author(s): Shigeishi H, Sugiyama M, Ohta K.
  • Journal Title: Biomed Rep Volume: 14 Pages: 40-40
  • Peer Reviewed
• [Journal Article] Detection of oral bacteria on the tongue dorsum using PCR amplification of 16S ribosomal RNA and its association with systemic disease in middle-aged and elderly patients. (2019)
  • Author(s): Su CY, Shigeishi H, Nishimura R, Ohta K, Sugiyama M.
  • Journal Title: Biomed Rep, Volume: 10 Pages: 70-76
  • Peer Reviewed / Open Access
• [Journal Article] Effect of hydrogel stiffness on morphology and gene expression pattern of CD44high oral squamous cell carcinoma cells. (2019)
  • Author(s): Shigeishi H, Yokoyama S, Murodumi H, Sakuma M, Kato H, Higashikawa K, Takechi M, Ohta K, Sugiyama M.
  • Journal Title: Int J Clin Exp Pathol. Volume: 12 Pages: 2826-2836
  • Peer Reviewed / Open Access
• [Journal Article] High HPV16 E6 viral load in the oral cavity is associated with an increased number of bacteria: A preliminary study. (2018)
  • Author(s): Shigeishi H, Sugiyama M, Ohta K, Yokoyama S, Sakuma M, Murozumi H, Kato H, Takechi M.
  • Journal Title: Biomed Rep Volume: 8 Pages: 59-64
  • Peer Reviewed
• [Journal Article] Human papillomavirus type 16 (HPV16) DNA copy number in oral rinse samples from oral cavity cancer patients. (2018)
  • Author(s): Shigeishi H, Yokoyama S, Ohta K, Takechi M, Sugiyama M.
  • Journal Title: Translational Research in Oral Oncology Volume: 3 Pages: 1-5
  • Peer Reviewed
• [Presentation] HPV16 DNAと歯周病原細菌との関係. (2021)
  • Author(s): 重石 英生，中村 真梨子，兼保 佳乃，杉山 勝，太田 耕司.
  • Organizer: 第70回日本口腔衛生学会・総会
• [Presentation] 口腔のHPV16感染と歯周病原細菌との関係 (2020)
  • Author(s): 重石 英生, 中村 真梨子，杉山 勝，太田 耕司
  • Organizer: 第104回広島大学歯学会例会
• [Presentation] Association between oral HPV prevalence and smoking in people without oral cancer (2019)
  • Author(s): Hideo Shigeishi, Masaru Sugiyama, Kouji Ohta
  • Organizer: 15th Annual Conference of the International Society for the Prevention of Tobacco Induced Diseases
  • Int'l Joint Research
• [Presentation] 口腔のHPV16と口腔衛生状態および歯周病原細菌との関係 (2019)
  • Author(s): 重石 英生，杉山 勝，太田 耕司
  • Organizer: 第28回日本口腔感染症学会総会・学術大会
• [Presentation] The association between oral HPV16 infection and oral health status in Japanese female aged 60 years and over (2019)
  • Author(s): Hideo Shigeishi, Cheng-Yih Su, Yoshino Kaneyasu, Mari Matsumura, Mariko Nakamura, Momoko Ishikawa, Ayumi Saito, Kouji Ohta, Masaru Sugiyama
  • Organizer: 第52回広島大学歯学会総会
• [Presentation] 口腔HPV感染の危険因子およびHPV陽性口腔癌の分子生物学的特徴 (2018)
  • Author(s): 重石 英生, 太田 耕司，杉山 勝.
  • Organizer: 第67回日本口腔衛生学会・総会
• [Presentation] 口腔含嗽サンプルを用いたHuman Papillomavirus (HPV) DNAの陽性率の検討. (2018)
  • Author(s): 重石 英生、太田 耕司，東川 晃一郎，武知 正晃，杉山 勝.
  • Organizer: 第47回（公社）日本口腔外科学会中国四国支部学術集会