Development of the alkylating agent targeting intrahepatic cholangiocarcinoma -specific fusion gene

2019 Fiscal Year Final Research Report

• Project/Area Number: 16K10583
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Digestive surgery
• Research Institution: Nihon University
• Principal Investigator: TAKAGI Keiko 日本大学, 医学部, 研究医員 (20339328)
• Co-Investigator(Kenkyū-buntansha): 緑川 泰 日本大学, 医学部, 准教授 (10292905) ; 尾崎 俊文 千葉県がんセンター(研究所), 発がん研究グループ DNA損傷シグナル研究室, 室長 (40260252) ; 藤原 恭子 日本大学, 歯学部, 准教授 (40595708)
• Project Period (FY): 2016-04-01 – 2020-03-31
• Keywords: 肝内胆管癌 / 融合遺伝子 / PIポリアミド

Outline of Final Research Achievements

We aimed to develop chlorambucil-PI polyamide (ChB-PIP), which alkylates a fusion gene specific for intrahepatic cholangiocarcinoma (ICC) and kills only cells contain the fusion gene.　ChB-PIP was prepared by purification and analysis by high performance liquid chromatography (HPLC), and the synthesis of ChB-PIP that specifically binds to the fusion gene was confirmed by gel shift assay. Then, a transient expression strain was prepared for the ICC cell line. Next, in order to establish a stable strain, after transfection, selection was performed using G418 (Geneticin) to scale up the cells.In the fusion gene plasmid-introduced group, the gene expression level was higher than that in the control group, and it was confirmed that a stable strain was produced.

Free Research Field

消化器外科

Academic Significance and Societal Importance of the Research Achievements

配列特異的にDNAに結合する性質を持つピロール・イミダゾール・ポリアミド(PIP)を用いて、融合領域を認識するPIPにアルキル化剤クロラムブシル(ChB)を付加したFGFR2-BICC1融合遺伝子認識ChB-PIPの合成を成功させた。また、融合遺伝子2種のいずれかの存在が確認されているICC細胞株が樹立されていないことから、安定株の作製を成功させた。