Search of lymph node metastasis controlling genes using Lenti-CRISPR-infected cancer cell in mouse model

2019 Fiscal Year Final Research Report

• Project/Area Number: 17K10577
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Digestive surgery
• Research Institution: Akita University
• Principal Investigator: Motoyama Satoru 秋田大学, 医学系研究科, 教授 (60292372)
• Co-Investigator(Kenkyū-buntansha): 佐藤 雄亮 秋田大学, 医学系研究科, 講師 (10431628)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: リンパ節転移モデル / in vivo passage / RNA-Seq / RNAメチル化

Outline of Final Research Achievements

The parental NR-S1M murine oral squamous cell carcinoma cells were serially in vivo passaged. Then, highly lymph node-metastatic and non-metastatic cell lines were derived from the same parental cells, and a reliable animal model of lymph node metastasis was established. RNA was extracted from both cell lines, and comprehensive gene expression analysis by RNA-Seq was performed to identify candidate factor X for controlling lymph node metastasis. We examined the correlation between the obtained candidate factor X and lymph node metastasis in esophageal cancer patients. In addition, we focused on RNA methylation N6-methyladenosine (m6A) and analyzed the effect of m6A demethylase on lymph node metastasis. The m6A demethylase regulates the growth of cancer cells and cell cycle analysis revealed that it is essential for the progression from G1 to S phase.

Free Research Field

消化器外科学

Academic Significance and Societal Importance of the Research Achievements

固形癌リンパ節転移に対する新しい制御法の開発が社会から切望されている。そこで、我々は全ての遺伝子に対するガイドRNAがプールされたPooled lenti-CRISPR libraryを感染させることによってリンパ節転移に関わる遺伝子を探索した。同時に、移植―リンパ節転移を繰り返すIn vivo passageよって今後の研究発展に不可欠なリンパ節転移動物モデルを確立した。さらにRNAメチル化のN6-メチルアデノシン(m6A)に着目し、m6A脱メチル化酵素のリンパ節転移に与える影響について研究した。これらの結果は今後のリンパ節転移研究において重要な知見を含んでおり、社会的意義が大きい。