Proteomic analysis identifies differentially expressed proteins after red propolis treatment in Hep-2 cells.
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 μg/mL (IC50): GRP78, PRDX2, LDHB, VIM and TUBA1A. Only two up-regulated proteins were identified in this study in the non-cytotoxic (6 μg/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 μg/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.
Laboratory of Genomics, Proteomics and DNA Repair, Biotechnology Institute, University of Caxias do Sul, RS, Brazil.
Caroline Olivieri da Silva
CO
Frozza
Laboratory of Genomics, Proteomics and DNA Repair, Biotechnology Institute, University of Caxias do Sul, RS, Brazil.
Tanara da Silva
Tda S
Ribeiro
Laboratory of Genomics, Proteomics and DNA Repair, Biotechnology Institute, University of Caxias do Sul, RS, Brazil.
Gabriela
G
Gambato
Laboratory of Genomics, Proteomics and DNA Repair, Biotechnology Institute, University of Caxias do Sul, RS, Brazil.
Caroline
C
Menti
Laboratory of Biotechnology of Natural and Synthetics Products, Biotechnology Institute, University of Caxias do Sul, RS, Brazil.
Sidnei
S
Moura
Laboratory of Applied Proteomics Campus São Gabriel, Federal University of Pampa, São Gabriel, RS, Brazil.
Paulo Marcos
PM
Pinto
Biotechnology Center, Federal University of Rio Grande do Sul, Av. Bento Gonçalves, Porto Alegre, RS, Brazil.
Charley Christian
CC
Staats
Program on Health and Environment, Tiradentes University, Aracaju, SE, Brazil.
Francine Ferreira
FF
Padilha
Biotechnology Unit, Center for Technology Development, Federal University of Pelotas, RS, Brazil.
Karine Rech
KR
Begnini
Biotechnology Unit, Center for Technology Development, Federal University of Pelotas, RS, Brazil.
Priscila Marques Moura
PM
de Leon
Biotechnology Unit, Center for Technology Development, Federal University of Pelotas, RS, Brazil.
Sibele
S
Borsuk
Biotechnology Unit, Center for Technology Development, Federal University of Pelotas, RS, Brazil.
Lucielli
L
Savegnago
Biotechnology Unit, Center for Technology Development, Federal University of Pelotas, RS, Brazil.
Odir
O
Dellagostin
Biotechnology Unit, Center for Technology Development, Federal University of Pelotas, RS, Brazil.
Tiago
T
Collares
Biotechnology Unit, Center for Technology Development, Federal University of Pelotas, RS, Brazil.
Fabiana Kömmling
FK
Seixas
Laboratory of Genomics, Proteomics and DNA Repair, Biotechnology Institute, University of Caxias do Sul, RS, Brazil.
João Antonio Pêgas
JA
Henriques
Laboratory of Genomics, Proteomics and DNA Repair, Biotechnology Institute, University of Caxias do Sul, RS, Brazil. Electronic address: mrely@ucs.br.
Mariana
M
Roesch-Ely
D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D000091342:Endoplasmic Reticulum Chaperone BiP; D005434:Flow Cytometry; D006801:Humans; D020287:In Situ Nick-End Labeling; D009363:Neoplasm Proteins / Q000378:metabolism*; D011429:Propolis / Q000494:pharmacology*; D040901:Proteomics; D053719:Tandem Mass Spectrometry
D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D009363:Neoplasm Proteins; D011429:Propolis
Annexin-V; Apoptosis; Hep-2; Proteomics; Red propolis; TUNEL