Establishment of a rapid intraoperative molecular analysis using the membrane patch method.

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K18728
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 56050:Otorhinolaryngology-related
• Research Institution: Hamamatsu University School of Medicine
• Principal Investigator: Yamaguchi Yuki 浜松医科大学, 医学部附属病院, 診療助教 (60837821)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: メンブレンパッチ法 / リキッドバイオプシー / エピジェネティクス解析

Outline of Final Research Achievements

In the first year, DNA was collected in three cases of tongue cancer using the membrane patch method; the average amount of DNA was 1.91 ug/ml on the tumor side and 1.19 ug/ml on the normal side. In the next year, the cytobrush method was introduced as an improvement to the membrane patch method. DNA was collected in 8 cases of oropharyngeal and tongue cancer cases. We were able to obtain DNA that were not different from the membrane patch method. In the final year, bisulfite treated DNA was analyzed the methylation status of E-cadherin, GALR1, TAC1, and COL1A2 was evaluated by Q-MSP method. Finally, we found that rapid molecular biological analysis by the cytobrush method was feasible within the same time frame as intraoperative pathology.

Free Research Field

頭頸部癌

Academic Significance and Societal Importance of the Research Achievements

切除後の迅速病理で断端に癌の残存を認めないにもかかわらず局所再発する、すなわち、顕微鏡的に確認できない癌や異形成の存在が疑われ、分子生物学的手法による切除断端、深部断端の癌残存有無の評価が必要である。
本申請は中咽頭癌、舌癌症例の切除後の深部断端における残存癌組織の有無を病理学的手法のみに頼らず、『メンブレンパッチ法』によるDNA採取と『サイトブラシ法』によるDNA採取を比較し、術中短時間HPV解析とDNAメチル化解析の実現を施行し、分子生物学的迅速解析法の確立のための基礎でデーター蓄積を行い、臨床へのリアルタイムな還元の可能性を証明した。