Establishment of a novel gene promoter activity detection system for cell isolation and characterization.

2015 Fiscal Year Final Research Report

• Project/Area Number: 26670815
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Pathobiological dentistry/Dental radiology
• Research Institution: Okayama University
• Principal Investigator: KOZAKI Ken-ichi 岡山大学, 大学院医歯薬学総合研究科, 教授 (50270715)
• Co-Investigator(Kenkyū-buntansha): SOGAWA Norio 松本歯科大学, 歯学部, 教授 (30236153) ; SOGAWA Chiharu 岡山大学, 大学院医歯薬学総合研究科, 講師 (10253022)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: 発現制御 / 遺伝子 / 癌 / 発生・分化 / 移植・再生医療

Outline of Final Research Achievements

We established a novel gene promoter activity detection (gPAD) system and utilized it in a cell-based drug screening, in which promoter sequences of marker genes were connected with fluorescent reporter genes and inserted in stable cell clones. Concretely, we established a gPAD system with a promoter sequence of VIM gene that encodes mesenchymal marker vimentin in order to evaluate alteration of epithelial-mesenchymal transition (EMT) program in cancer cells and then performed functional screening using the cell-based reporter system. We established such VIM gPAD system within MKN1 colon cancer cells, performed screening from 328 species of miRNAs, and then identified a novel EMT-inducing microRNA miR-544a. miR-544a was shown to promote EMT through targeting CDH1 gene, encoding an epithelial marker E-cadherin, and targeting AXIN2 gene, encoding a Wnt signaling inhibitory factor, and thus to activate Wnt and TGFβ signaling pathways (Yanaka Y et al., 2015, Carcinogenesis). As above, an example and usefulness of gPAD system in drug screening were shown.

Free Research Field

医歯薬学