| Project/Area Number |
24650620
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Tumor biology
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| Research Institution | Kyoto University |
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Principal Investigator |
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| Project Period (FY) |
2012-04-01 – 2014-03-31
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2012: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | EMT / 乳癌幹細胞 / MET / レポーター / 発現クローニング / トリプルネガティブ乳癌 / 上皮間葉転換 (EMT) / 間葉上皮転換 (MET) / 乳癌 / ライブラリースクリーニング |
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| Research Abstract |
In order to identify cDNA/shRNA that can induce EMT or MET, we established a reporter system where vimentin or E-cadherin promoter drives blasticidine-resistant gene and GFP in HMLE epithelial or MDA-MB-231 mesenchymal breast cancer cell line. The cells were infected with cDNA or shRNA library and subjected to blasticidine selection. In EMT screening, Snail and Twist2 were identified as EMT inducer, and two genes, members of mitochondrial complex I, and a hub gene in signal transduction were identified to support soft aga-colonogenecity of mesenchymal cells. In MET screen, a potent MET-inducing shRNA (called shP1) was identified. The shP1 was tested for soft agar colony formation using monoclonal dual reporter cell clones from SUM159 mesenchymal breast cancer cell line. Interestingly, there are two sets of reporter clones with different response to shP1. Colony formation did not decreased in one set of reporter clones, while in the other clone shP1 reduced colony formation.
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