A dietary polyacetylene, Gymnasterkoreayne B, induces both intrinsic and extrinsic pathway of apoptosis through the signaling molecules revealed by secretome profiling
- A dietary polyacetylene, Gymnasterkoreayne B, induces both intrinsic and extrinsic pathway of apoptosis through the signaling molecules revealed by secretome profiling
- 송대근; 강경수; 이은하; 정상훈; 판철호; 노주원
- secretome; proteomics; gymnasterkoreayne b; gymnaster koraiensis; apoptosis; hct116; intrinsic; extrinsic; lc-ms; facs
- Issue Date
- Experimental biology 2014
- Gymnasterkoreayne B (GKB) is a natural polyacetylene isolated from Gymnaster koraiensis. The antiproliferative activity of GKB has been reported in various human cancer cells but its mechanism is still unknown. Our objectivity of the study was to investigate the mechanism of the antiproliferative activity of GKB in HCT116 cancer cell and to search any possible candidates that may involve in the cell death signaling.
In this study, we confirmed that GKB has induced apoptosis in HCT116 cells, inferentially from the increment of the sub G0/G1 content (%) and the externalization of phosphatidylserine, and the apoptotic morphological changes. We also verified that the antiproliferative activity of GKB was not necroptosis, by the cell death inhibitor assay using z-VAD-fmk and necrostatin. In addition, by the mitochondrial membrane potential assay and flow-cytometric analysis, we elucidated that GKB treatment resulted in ROS accumulation, leading to the activation of both intrinsic and extrinsic apoptotic pathway. To investigate the effect of GKB on the secretome profile, we compared protein levels in the media of GKB- or vehicle-treated HCT116 cells using 1D-SDS-PAGE and LC-MS/MS analysis. Total of 1804 proteins were identified and their expression levels were determined by using spectral counting method. As a result, 70 proteins were down regulated (< 0.5 fold) and 56 proteins were up regulated (> 2.0 fold). Among these, the expression levels of apoptosis-related proteins, ADAM9, interleukin 18, and serpin B5 were validated by ELISA. In depth analysis regarding the gene ontology of the identified protein suggested that FN1, TGFB1, APP, SERPINE1, HSPD1, SOD1, TXN, and ACTN4 may also act as secretory signaling molecules that trigger GKB-induced apoptosis.
In summary, we have found the mechanism of GKB-induced apoptosis of HCT116 and we have suggested novel proteins which might be involved in the apoptotic signaling.
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