Secretome Analysis of Gymnasterkoreayne B Treated HCT116 Cells

Secretome Analysis of Gymnasterkoreayne B Treated HCT116 Cells
secretome; lc-ms; Gymnater koraiensis; Gymnaterkoreayne B; apoptosis; HCT116; ELISA; spectrum counting; GO term
Issue Date
한국질량분석학회 2012여름정기학술대회
Gymnasterkoreayne B (GKB) is a cancer chemopreventive polyacetylene, isolated from a Korean wild vegetable, Gymnaster koraiensis. When GKB was treated at higher dose, HCT116 cells underwent apoptosis, suggesting that GKB may serve as an anti-cancer agent. Recently, there are attempts to explore the crosstalk between the apoptotic cells by studying secretome. In this study, we investigated the mechanism of apoptosis induced by GKB in HCT116 cells, in terms of the influence on the secretome profile. We compared the protein levels in the media of GKB treated and un-treated HCT116 cells using 1D-SDS-PAGE followed by LC-MS/MS analysis. Of the secreted proteins, which were filtered by using GO terms and DAVID tool, the expression levels were determined by Spectral Counting. Additionally, student’s t-test was applied and the expression levels of some proteins were confirmed by ELISA. As a result, total of 1804 proteins were identified. After the filtering process, 80 proteins were down regulated (>2.0 fold) and 58 proteins were up regulated (<0.5 fold). The expression level of ADAM9 (down), interleukin 18 (IL18) and serpin B5 (up) were confirmed by ELISA. Our study may serve as a foothold for studying the crosstalk between the GKB-induced apoptotic cells.
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