Proteomic analysis identifies differentially expressed proteins after red propolis treatment in Hep-2 cells
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- 作者：Caroline Olivieri da Silva Frozza ; Tanara da Silva Ribeiro ; Gabriela Gambato ; Caroline Menti ; Sidnei Moura ; Paulo Marcos Pinto ; Charley Christian Staats ; Francine Ferreira Padilha ; Karine Rech Begnini ; Priscila Marques Moura de Leon ; Sibele Borsuk ; Lucielli Savegnago ; Odir Dellagostin ; Tiago Collares ; Fabiana K枚mmling Seixas ; Jo茫o Antonio P锚gas Henriques ; Mariana Roesch-Ely
- 关键词：Red propolis ; Hep-2 ; Proteomics ; Apoptosis ; TUNEL ; Annexin-V
- 刊名：Food and Chemical Toxicology
- 出版年：January, 2014
- 年：2014
- 卷：63
- 期：Complete
- 页码：195-204
- 全文大小：1139 K
文摘
Here we investigated alterations in the protein profile of Hep-2 treated with red propolis using two-dimensional electrophoresis associated to mass spectrometry and apoptotic rates of cells treated with and without red propolis extracts through TUNEL and Annexin-V assays. A total of 325 spots were manually excised from the two-dimensional gel electrophoresis and 177 proteins were identified using LC-MS-MS. Among all proteins identified that presented differential expression, most were down-regulated in presence of red propolis extract at a concentration of 120 渭g/mL (IC50): GRP78, PRDX2, LDHB, VIM and TUBA1A. Only two up-regulated proteins were identified in this study in the non-cytotoxic (6 渭g/mL) red propolis treated group: RPLP0 and RAD23B. TUNEL staining assay showed a markedly increase in the mid- to late-stage apoptosis of Hep-2 cells induced by red propolis at concentrations of 60 and 120 渭g/mL when compared with non-treated cells. The increase of late apoptosis was confirmed by in situ Annexin-V analysis in which red propolis extract induced late apoptosis in a dose-dependent manner. The differences in tumor cell protein profiles warrant further investigations including isolation of major bioactive compounds of red propolis in different cell lines using proteomics and molecular tests to validate the protein expression here observed.