Brazilian red propolis extracts: study of chemical composition by ESI-MS/MS (ESI+) and cytotoxic profiles against colon cancer cell lines
Denis Amiltondos Santos, Fernanda Mosena Munari, Caroline Olivieri da Silva Frozza, Sidnei Moura, Thiago Barcellos, João Antonio Pêgas Henriques, Mariana Roesch-Ely
Abstract
Red propolis is a natural resin mixture produced by honeybees and presents a source of active compounds with a variety of biological activities. In this study, we describe the chemical characterization and potential antitumor activity of total extract of Brazilian red propolis and its fractions. Fractions were obtained through column chromatography revealing 14 different compounds in all samples, which were determined and distinguished of other isobar molecules by fragmentation pathways by ESI-MS/MS in positive mode. Some molecules as cis-asarone or trans-isoelemicin were identified and distinguish from elemicin compound and vestitol or isovestitol were also distinguished from neovestitol by fragmention pathway. Other important compounds as liquiritigenin was differentiated from isoliquiritigenin and formononetin from dalbergin.
MTT viability assay showed different toxicity in cell lines after exposition to total extract and fractions. Fractions 05 and 06 had more selectivity against HT-29 and HCT-116 cancer cells, respectively, in relation to normal cells. IC50 (ranging of 72.45 ± 6.57 to 73.58 ± 1.00 μg/mL) in cancer cells were lower than reported in total extracts of propolis. May-Grunwald/Giemsa staining revealed cellular morphological changes after exposition to higher concentrations of red propolis extracts. Fractionation techniques can contribute to reduce chemical diversity verified in propolis mixtures, generating fractions with improved biological activity and contributing to the development of new strategies for discovery of natural compounds against cancer.
Keywords
References
Alencar et al., 2007
S.M. Alencar, T.L.C. Oldoni, M.L. Castro, I.S.R. Cabral, C.M. Costa-Neto, J.A. Cury, et al.
Chemical composition and biological activity of a new type of Brazilian propolis: Red propolis
Journal of Ethnopharmacology, 113 (2) (2007), pp. 278-283, 10.1016/j.jep.2007.06.005
Awale et al., 2008
S. Awale, F. Li, H. Onozuka, H. Esumi, Y. Tezuka, S. Kadota
Constituents of Brazilian red propolis and their preferential cytotoxic activity against human pancreatic PANC-1 cancer cell line in nutrient-deprived condition
Bioorganic and Medicinal Chemistry, 16 (1) (2008), pp. 181-189, 10.1016/j.bmc.2007.10.004
Bankova, 2005
V. Bankova
Chemical diversity of propolis and the problem of standardization
Journal of Ethnopharmacology, 100 (2005), pp. 114-117, 10.1016/j.jep.2005.05.004
Bankova et al., 2014
V. Bankova, M. Popova, B. Trusheva
Propolis volatile compounds: Chemical diversity and biological activity: A review
Chemistry Central Journal (2014), 10.1186/1752-153X-8-28
Barud et al., 2013
H.D.S. Barud, A.M. De Araújo Júnior, S. Saska, L.B. Mestieri, J.A.D.B. Campos, R.M. De Freitas, et al.
Antimicrobial Brazilian propolis (EPP-AF) containing biocellulose membranes as promising biomaterial for skin wound healing
Evidence-Based Complementary and Alternative Medicine, 2013 (2013), 10.1155/2013/703024
Begnini et al., 2014
K.R. Begnini, P.M. Moura De Leon, H. Thurow, E. Schultze, V.F. Campos, F. Martins Rodrigues, et al.
Brazilian red propolis induces apoptosis-like cell death and decreases migration potential in bladder cancer cells
Evidence-Based Complementary and Alternative Medicine, 2014 (2014), 10.1155/2014/639856
Boudet, 2007
A.M. Boudet
Evolution and current status of research in phenolic compounds
Phytochemistry, 68 (2007), pp. 2722-2735, 10.1016/j.phytochem.2007.06.012
Catchpole et al., 2015
O. Catchpole, K. Mitchell, S. Bloor, P. Davis, A. Suddes
Antiproliferative activity of New Zealand propolis and phenolic compounds vs human colorectal adenocarcinoma cells
Fitoterapia, 106 (2015), pp. 167-174, 10.1016/j.fitote.2015.09.004
Choi et al., 2013
R.C.Y. Choi, J.T.T. Zhu, A.W.Y. Yung, P.S.C. Lee, S.L. Xu, A.J.Y. Guo, et al.
Synergistic action of flavonoids, baicalein, and daidzein in estrogenic and neuroprotective effects: A development of potential health products and therapeutic drugs against alzheimer's disease
Evidence-Based Complementary and Alternative Medicine (2013), 10.1155/2013/635694
Choudhari et al., 2013
M.K. Choudhari, R. Haghniaz, J.M. Rajwade, K.M. Paknikar
Anticancer activity of Indian stingless bee propolis: An in vitro study
Evidence-Based Complementary and Alternative Medicine (2013), 10.1155/2013/928280
Cuesta-Rubio et al., 2007
O. Cuesta-Rubio, A.L. Piccinelli, M.C. Fernandez, I.M. Hernández, A. Rosado, L. Rastrelli
Chemical characterization of Cuban propolis by HPLC-PDA, HPLC-MS, and NMR: The brown, red, and yellow Cuban varieties of propolis
Journal of Agricultural and Food Chemistry (2007), 10.1021/jf071296w
Daugsch et al., 2008
A. Daugsch, C.S. Moraes, P. Fort, Y.K. Park
Brazilian red propolis – Chemical composition and botanical origin
Evidence-Based Complementary and Alternative Medicine (2008), 10.1093/ecam/nem057
Elbaz et al., 2016
N.M. Elbaz, I.A. Khalil, A.A. Abd-Rabou, I.M. El-Sherbiny
Chitosan-based nano-in-microparticle carriers for enhanced oral delivery and anticancer activity of propolis
International Journal of Biological Macromolecules, 92 (2016), 10.1016/j.ijbiomac.2016.07.024
Elsevier B.V.
Fasolo et al., 2016
D. Fasolo, A.M. Bergold, G. von Poser, H.F. Teixeira
Determination of benzophenones in lipophilic extract of Brazilian red propolis, nanotechnology-based product and porcine skin and mucosa: Analytical and bioanalytical assays
Journal of Pharmaceutical and Biomedical Analysis, 124 (2016), pp. 57-66, 10.1016/j.jpba.2016.02.018
Franchin et al., 2016
M. Franchin, D.F. Cólon, F.V.S. Castanheira, M.G. Da Cunha, B. Bueno-Silva, S.M. Alencar, et al.
Vestitol isolated from Brazilian Red Propolis inhibits neutrophils migration in the inflammatory process: Elucidation of the mechanism of action
Journal of Natural Products, 79 (4) (2016), pp. 954-960, 10.1021/acs.jnatprod.5b00938
Freires et al., 2016
I.A. Freires, S.M. de Alencar, P.L. Rosalen
A pharmacological perspective on the use of Brazilian Red Propolis and its isolated compounds against human diseases
European Journal of Medicinal Chemistry, 110 (2016), pp. 267-279, 10.1016/j.ejmech.2016.01.033
Frozza et al., 2013
C.O. Frozza, C.S.C. Garcia, G. Gambato, M.D.O. de Souza, M. Salvador, S. Moura, et al.
Chemical characterization, antioxidant and cytotoxic activities of Brazilian red propolis
Food and Chemical Toxicology, 52 (2013), pp. 137-142, 10.1016/j.fct.2012.11.013
Frozza et al., 2017
C.O. da S. Frozza, D.A. Santos, L.C. Rufatto, L. Minetto, F.J. Scariot, S. Echeverrigaray, et al.
Antitumor activity of Brazilian red propolis fractions against Hep-2 cancer cell line
Biomedicine and Pharmacotherapy (2017), 10.1016/j.biopha.2017.05.027
Harvey et al., 2015
A.L. Harvey, R. Edrada-Ebel, R.J. Quinn
The re-emergence of natural products for drug discovery in the genomics era
Nature Reviews Drug Discovery (2015), 10.1038/nrd4510
Hsu et al., 2018
Y.N. Hsu, H.W. Shyu, T.W. Hu, J.P. Yeh, Y.W. Lin, L.Y. Lee, et al.
Anti-proliferative activity of biochanin A in human osteosarcoma cells via mitochondrial-involved apoptosis
Food and Chemical Toxicology, 112 (December 2017) (2018), pp. 194-204, 10.1016/j.fct.2017.12.062
Huang et al., 2014
S. Huang, C.P. Zhang, K. Wang, G.Q. Li, F.L. Hu
Recent advances in the chemical composition of propolis
Molecules, 19 (12) (2014), pp. 19610-19632, 10.3390/molecules191219610
Huang et al., 2015
J. Huang, M. Xie, P. Gao, Y. Ye, Y. Liu, Y. Zhao, et al.
Antiproliferative effects of formononetin on human colorectal cancer via suppressing cell growth in vitro and in vivo
Process Biochemistry, 50 (6) (2015), pp. 912-917, 10.1016/j.procbio.2015.03.001
INCA, 2016
Instituto Nacional de Câncer José Alencar Gomes da Silva
INCA – Instituto Nacional de Câncer – Estimativa 2016
Ministério da Saúde Instituto Nacional de Cancer José Alencar Gomes da Silva (2016)
DOI: 978-85-7318-283-5
Kamiya et al., 2012
T. Kamiya, H. Nishihara, H. Hara, T. Adachi
Ethanol extract of Brazilian red propolis induces apoptosis in human breast cancer MCF-7 cells through endoplasmic reticulum stress
Journal of Agricultural and Food Chemistry (2012), 10.1021/jf303004n
Kuete et al., 2013
V. Kuete, P.D. Tchakam, B. Wiench, B. Ngameni, H.K. Wabo, M.F. Tala, et al.
Cytotoxicity and modes of action of four naturally occuring benzophenones: 2,2′,5,6′-Tetrahydroxybenzophenone, guttiferone E, isogarcinol and isoxanthochymol
Phytomedicine (2013), 10.1016/j.phymed.2013.02.003
Kuropatnicki et al., 2013
A.K. Kuropatnicki, E. Szliszka, W. Krol
Historical aspects of propolis research in modern times
Evidence-Based Complementary and Alternative Medicine, 2013 (2013), 10.1155/2013/964149
Lahouel et al., 2007
M. Lahouel, S. Amedah, A. Zellagui, A. Touil, S. Rhouati, F. Benyache, et al.
The interaction of new plant flavonoids with rat liver mitochondria: Relation between the anti- and pro-oxydant effect and flavonoids concentration
Thérapie (2007), 10.2515/therapie:2006025
Li et al., 2008
F. Li, S. Awale, Y. Tezuka, S. Kadota
Cytotoxic constituents from Brazilian red propolis and their structure-activity relationship
Bioorganic and Medicinal Chemistry, 16 (10) (2008), pp. 5434-5440, 10.1016/j.bmc.2008.04.016
Li et al., 2014
T. Li, X. Zhao, Z. Mo, W. Huang, H. Yan, Z. Ling, et al.
Formononetin promotes cell cycle arrest via downregulation of akt/cyclin D1/CDK4 in human prostate cancer cells
Cellular Physiology and Biochemistry, 34 (4) (2014), pp. 1351-1358, 10.1159/000366342
Liu et al., 2011
C. Liu, Y. Wang, S. Xie, Y. Zhou, X. Ren, X. Li, et al.
Liquiritigenin induces mitochondria-mediated apoptosis via cytochrome c release and caspases activation in heLa cells
Phytotherapy Research, 25 (2) (2011), pp. 277-283, 10.1002/ptr.3259
Liu et al., 2012
Y. Liu, S. Xie, Y. Wang, K. Luo, Y. Wang, Y. Cai
Liquiritigenin inhibits tumor growth and vascularization in a mouse model of HeLa cells
Molecules, 17 (6) (2012), pp. 7206-7216, 10.3390/molecules17067206
Lopez et al., 2015
B.G.C. Lopez, C.C. de Lourenço, D.A. Alves, D. Machado, M. Lancellotti, A.C.H.F. Sawaya
Antimicrobial and cytotoxic activity of red propolis: An alert for its safe use
Journal of Applied Microbiology, 119 (3) (2015), pp. 677-687, 10.1111/jam.12874
López et al., 2014
B.G.C. López, E.M. Schmidt, M.N. Eberlin, A.C.H.F. Sawaya
Phytochemical markers of different types of red propolis
Food Chemistry, 146 (2014), pp. 174-180, 10.1016/j.foodchem.2013.09.063
Mendonça et al., 2015
I.C.G. Mendonça, I.C.C. de M. Porto, T.G. do Nascimento, N.S. de Souza, J.M. dos S. Oliveira, R.E. dos S. Arruda, et al.
Brazilian red propolis: Phytochemical screening, antioxidant activity and effect against cancer cells
BMC Complementary and Alternative Medicine (2015), 10.1111/j.1757-5672.2011.00097.x
Mosmann, 1983
T. Mosmann
Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays
Journal of Immunological Methods (1983), 10.1016/0022-1759(83)90303-4
Mouhoubi-Tafinine et al., 2016
Z. Mouhoubi-Tafinine, S. Ouchemoukh, A. Tamendjari
Antioxydant activity of some algerian honey and propolis
Industrial Crops and Products (2016), 10.1016/j.indcrop.2016.02.033
Najafi et al., 2007
M.F. Najafi, F. Vahedy, M. Seyyedin, H.R. Jomehzadeh, K. Bozary
Effect of the water extracts of propolis on stimulation and inhibition of different cells
Cytotechnology, 54 (1) (2007), pp. 49-56, 10.1007/s10616-007-9067-2
Novak et al., 2014
E.M. Novak, M.S.C. Silva, M.C. Marcucci, A.C.H.F. Sawaya, B. Gimenez-Cassina Lopez, M.A.H. Z. Fortes, et al.
Antitumoural activity of Brazilian red propolis fraction enriched with xanthochymol and formononetin: An in vitro and in vivo study
Journal of Functional Foods, 11 (C) (2014), pp. 91-102, 10.1016/j.jff.2014.09.008
Nunes and Guerreiro, 2012
C.A. Nunes, M.C. Guerreiro
Characterization of Brazilian green propolis throughout the seasons by headspace GC/MS and ESI-MS
Journal of the Science of Food and Agriculture, 92 (2) (2012), pp. 433-438, 10.1002/jsfa.4596
Piccinelli et al., 2011
A.L. Piccinelli, C. Lotti, L. Campone, O. Cuesta-Rubio, M. Campo Fernandez, L. Rastrelli
Cuban and Brazilian red propolis: Botanical origin and comparative analysis by high-performance liquid chromatography-photodiode array detection/electrospray ionization tandem mass spectrometry
Journal of Agricultural and Food Chemistry, 59 (12) (2011), pp. 6484-6491, 10.1021/jf201280z
Protiva et al., 2008
P. Protiva, M.E. Hopkins, S. Baggett, H. Yang, M. Lipkin, P.R. Holt, et al.
Growth inhibition of colon cancer cells by polyisoprenylated benzophenones is associated with induction of the endoplasmic reticulum response
International Journal of Cancer, 123 (3) (2008), pp. 687-694, 10.1002/ijc.23515
Puthli et al., 2013
A. Puthli, R. Tiwari, K.P. Mishra
Biochanin A enhances the radiotoxicity in colon tumor cells in vitro
Journal of Environmental Pathology, Toxicology and Oncology (2013), 10.1615/JEnvironPatholToxicolOncol.2013007280
Raheja et al., 2018
S. Raheja, A. Girdhar, V. Lather, D. Pandita
Biochanin A: A phytoestrogen with therapeutic potential
Trends in Food Science and Technology, 79 (2018), pp. 55-66, 10.1016/j.tifs.2018.07.001
Righi et al., 2011
A.A. Righi, T.R. Alves, G. Negri, L.M. Marques, H. Breyer, A. Salatino
Brazilian red propolis: Unreported substances, antioxidant and antimicrobial activities
Journal of the Science of Food and Agriculture, 91 (13) (2011), pp. 2363-2370, 10.1002/jsfa.4468
Roleira et al., 2015
F.M.F. Roleira, E.J. Tavares-Da-Silva, C.L. Varela, S.C. Costa, T. Silva, J. Garrido, et al.
Plant derived and dietary phenolic antioxidants: Anticancer properties
Food Chemistry, 183 (2015), pp. 235-258, 10.1016/j.foodchem.2015.03.039
Rufatto et al., 2017
L.C. Rufatto, D.A. dos Santos, F. Marinho, J.A.P. Henriques, M. Roesch Ely, S. Moura
Red propolis: Chemical composition and pharmacological activity
Asian Pacific Journal of Tropical Biomedicine (2017), 10.1016/j.apjtb.2017.06.009
Siegel et al., 2016
R.L.R.L. Siegel, D.K. Miller, A. Jemal
Cancer Statistics (2016), 10.3322/caac.21332
Silva et al., 2008
B.B. Silva, P.L. Rosalen, J.A. Cury, M. Ikegaki, V.C. Souza, A. Esteves, et al.
Chemical composition and botanical origin of red propolis, a new type of Brazilian propolis
Evidence-Based Complementary and Alternative Medicine, 5 (3) (2008), pp. 313-316, 10.1016/j.clay.2018.10.007
Silva-Carvalho et al., 2015
R. Silva-Carvalho, F. Baltazar, C. Almeida-Aguiar
Propolis: A complex natural product with a plethora of biological activities that can be explored for drug development
Evidence-Based Complementary and Alternative Medicine, 2015 (2015), 10.1155/2015/206439
Toreti et al., 2013
V.C. Toreti, H.H. Sato, G.M. Pastore, Y.K. Park
Recent progress of propolis for its biological and chemical compositions and its botanical origin
Evidence-Based Complementary and Alternative Medicine (2013), 10.1155/2013/697390
Trusheva et al., 2006
B. Trusheva, M. Popova, V. Bankova, S. Simova, M.C. Marcucci, P.L. Miorin, et al.
Bioactive constituents of Brazilian red propolis
Evidence-Based Complementary and Alternative Medicine, 3 (2) (2006), pp. 249-254, 10.1093/ecam/nel006
Wagh, 2013
V.D. Wagh
Propolis: A wonder bees product and its pharmacological potentials
Advances in Pharmacological Sciences (2013), 10.1155/2013/308249
Watanabe et al., 2011
M.A.E. Watanabe, M.K. Amarante, B.J. Conti, J.M. Sforcin
Cytotoxic constituents of propolis inducing anticancer effects: A review
The Journal of Pharmacy and Pharmacology, 63 (11) (2011), pp. 1378-1386, 10.1111/j.2042-7158.2011.01331.x
Wu et al., 2015
X.Y. Wu, H. Xu, Z.F. Wu, C. Chen, J.Y. Liu, G.N. Wu, et al.
Formononetin, a novel FGFR2 inhibitor, potently inhibits angiogenesis and tumor growth in preclinical models
Oncotarget (2015), 10.18632/oncotarget.6310
Zancanela et al., 2018
D.C. Zancanela, C.S. Funari, R.D. Herculano, V.M. Mello, C.M. Rodrigues, F.A. Borges, et al.
Natural rubber latex membranes incorporated with three different types of propolis: Physical-chemistry and antimicrobial behaviours
Materials Science and Engineering (2018), 10.1016/j.msec.2018.12.042