2015 Fiscal Year Final Research Report
Intracellular kinetic analysis of functional food component by CLEM method
| Project/Area Number |
25450181
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Food science
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| Research Institution | Tokyo University of Agriculture |
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Principal Investigator |
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| Project Period (FY) |
2013-04-01 – 2016-03-31
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| Keywords | ポリフェノール / オリーブ葉 / 蛍光標識 / CLEM法 / 機能性食品 / オレウロペイン / ヒドロキシチロソール / 大腸癌 |
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| Outline of Final Research Achievements |
The aim of this study was to investigate the novel beneficial effects of hydroxytyrosol (HT) using colorectal cancer (Caco-2) cells and to analyze the localization of HT conjugated to fluorescein isothiocyanate (FITC) in Caco-2 using Corrlative light and electron microscopy (CLEM) method.
Our results revealed that HT inhibited cell proliferation dose-dependently in Caco-2 cells and induced DNA fragmentation, suggesting that HT induced apoptotic cell death. Furthermore, HT inhibited Akt/mTOR pathway to induce autophagy. Furthermore, HT was conjugated with FITC via the alcoholic hydroxyl group (FITC-HT). Caco-2 cells were incubated with 0.5 mM FITC-HT for 24 hours and subsequently labelled with Lysosome marker before imaging on the Olympus FV1200 Laser confocal microscope. As a result, FITC-HT was detected in the lysosome of Caco-2. Additionally, CLEM was performed using Caco-2 cells treated with FITC-HT and LysoTracker. As a result, primitive imaging data was obtained.
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| Free Research Field |
栄養生化学
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