Comprehensive screening of genes resistant to Sorafenib in Hepatocellular carcinoma

Research Project

Project/Area Number	26461019
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Research Field	Gastroenterology
Research Institution	Keio University
Principal Investigator	Abe Yuta 慶應義塾大学, 医学部(信濃町), 講師 (70327526)
Co-Investigator(Renkei-kenkyūsha)	Hayashida Tetsu 慶應義塾大学, 医学部, 講師 (80327543)
Project Period (FY)	2014-04-01 – 2017-03-31
Project Status	Completed (Fiscal Year 2016)
Budget Amount *help	¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000) Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000) Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000) Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywords	肝細胞癌 / 化学療法 / ソラフェニブ / 放射線治療 / 薬剤耐性 / 放射線耐性
Outline of Final Research Achievements	We used a novel method involving transposons to screen and identify drug-resistant genes. Transposons are DNA sequences that move from one location on the gene to another. A modified piggyBac transposon was designed as an insertion mutagen, and a cytomegalovirus (CMV) promoter sequence was added to induce strong transcription. When the transposon is inserted to the upstream of a certain gene, the gene will be overexpressed while when intserted down or intragenically, it will be downregulated. After establishing a transposon-tagged cell library, we treated cell lines derived from Hepatocellular carcinoma. We performed splinkerette PCR and TOPO cloning on the resistant colonies. Bacterial colonies were sequenced, and next-generation sequencing was used to identify the overexpressed/downregulated sequences as candidate genes for Sorafenib resistance. We identified several candidate genes from 30 resistant colonies.