MODULATOR IMPACTS OF PROPOLIS EXTRACT AGAINST DOXORUBICIN MEDIATED CARCINOGENESIS ON HEPATOCELLULAR CARCINOMA AND Drosophila SOMATIC CELLS
Abstract
The antitumor action of propolis is of clinical interest because of the need for new anticancer treatment agents. The present investigation intended to extract and assess the chemical content, cytotoxic action, the growth inhibitory activity and anticancer capability of Egyptian propolis versus Chinese propolis. This was carried out using water extract (WE) and ethanolic extract (EE) on the human hepatocellular carcinoma (HEp-2) cell line and the loss of heterozygosity (LOH) assay of Drosophila melanogaster somatic cells against the direct genotoxicity of doxorubicin. EPWE, EPEE, CPWE and CPEE extracts analyzed by HPLC showed that there were sensible and various concentrations of phenolic compounds in both. Total phenolics were determined to be 18.83, 34.87, 39.29 and 180.89 g-1 by using EPWE, CPWE, EPEE and CPEE extracts, respectively. Chinese propolis ethanol extract (CPEE) have major concentrations of total phenolics and phenolic acids and contained high concentrations of rutin (188.90 g/mL). The study of the antiproliferative capacity of propolis extractors against HEp-2 cancer cell lines showed that all the studied propolis extracts induce suppression of cell growth except CPWE extract; it gave 100% cell viability. The great majority of the propolis are strongly cytotoxic against HEp-2 cell line with 500 μg/ml CPEE. Also, PEE is the most effective in inhibition of HEp-2 cell proliferation compared to PWE. In Drosophila assay, treatment with propolis extract and DOX carcinogenic agent led to a reduction in the frequency of recombination compared to the treatment with DOX alone either in the post- and pre-treatments. In general, PEE exhibited powerful anti-proliferative effects than PWE. The ethanol extract provided the highest protection against Doxorubicin (DOR) induced genotoxicity, a fact that supports their anti-cancer activity. The results demonstrate that PEE is a good source of a natural antitumor operator able to inhibit cancer cell proliferation.References
Ahn, M. R., K. Kazuhiro, K. Shigenori, N. Tsutomu, K. Kazuhiko, U. Yoshihiro, H. Hitoshi, N. Hideko and O. Toshiro (2009). Correlation between antiangiogenic activity and antioxidant activity of various components from propolis. Molecular Nutrition & Food Research, 53: 643-651.
Bankova, V. S., S. L. De Castro and M. C. Marcucci (2000). Propolis: recent advances in chemistry and plant origin. Apidologie, 31: 3-15. Banskota, A. H., Y. Tezuka and S. H. Kadota (2001). Recent progress in pharmacological research of propolis. Phytotherapy Research, 15: 561-571.
Barlak, Y., O. Deger, M. Colak, S. C. Karatayli, A. M. Bozdayi and F. Yucesan (2011). Effect of Turkish propolis extracts on proteome of prostate cancer cell line. Proteome Sci., 9: 74.
Bissery, M. C., F. A. Valeriote, G. G. Chabot, J. D. Crisman, C. Yost and T. H Corbett (1988). Flavone acetic acid (NSC-34 7512)-induced DNA damage in Glasgow osteogenic sarcoma in vivo. Cancer Research, 48: 1279-1285.
Borges, J., M. Tagliamento, A. Silva, P. Sobral and R. Carvalho (2013). Development and characterization of orally-disintegrating films for propolis delivery. Food Science and Technology, Ciênc. Tecnol. Aliment., 33: 28-33.
Borges, K. S., M. S. Brassesco, C. A. Scrideli, A. E. Soares and L. G. Tone (2011). Anti-proliferative effects of Tubi-bee propolis in glioblastoma cell lines 2. Genet. Mol. Biol., 34: 310-314.
Burdock, G. A. (1998). Review of the biological properties and toxicity of bee propolis, Food Chem. Toxicol., 36: 347-363.
Chen, M. J., W. H. Chang, C. C. Lin, C. Y. Liu, T. E. Wang, C. H. Chu, S. C. Shih and Y. J. Chen (2008). Caffeic acid phenethyl ester induces apoptosis of human pancreatic cancer cells involving caspase and mitochondrial dysfunction. Pancreatology, 8: 566-76.
Chirumbolo, S. (2012). Flavonoids in propolis acting on mast cellmediated wound healing. Inflammopharmacology, 20: 99- 101.
Christov, R., B. Trusheva, M. Popova, V. Bankova and M. Bertrand (2005). Chemical composition of propolis from Canada, its antiradical activity and plant origin. Natural Product Research, 19: 673-678
Crane, E. (1997). The past and present importance of bee products to man. Bee Products: Properties, Applications and Apitherapy. Plenum, New York.
Cunha, I. B. S., A. C. H. F. Sawaya, F. M. Caetano, M. T. Shimizu, M. C. Marcucci, F. T. Drezza, G. S. Povia and P. O. Carvalho (2004). Factors that influence the yield and composition of Brazilian propolis extracts. Journal of the Brazilian Chemical Society, 15: 964-970.
Czepiel, J., G. Biesiada, M. Gajda, W. Szczepan´ski, K. Szypuła, Z. Dabrowski and T. Mach (2010). The effect of TCDD dioxin on the rat liver in biochemical and histological assessment. Folia Biologica (Krakow), 58: 85-90.
Da Silva, F. C. O., G. C. S. Celi, G. Gambato, M. D. Oliveira de Souza, M. Salvador, S. Moura, F. F. Padilha, F. K. Seixas, T. Collares, S. Borsuk, O. A. Dellagostin, J. A. P. Henriques and M. Roesch-Ely (2013). Chemical characterization, antioxidant and cytotoxic activities of Brazilian red propolis. Food Chem. Toxicol., 52: 137-142.
Daugsch, A., C. S. Moraes, P. Fort and Y. K. Park (2008). Brazilian red propolis - chemical composition and botanical origin. Evid Based Complement Alternat. Med., 5: 435-441.
Díaz-Carballo, D., S. Malak, W. Bardenheuer, M. Freistuehler and H. P. Reusch (2008). The contribution of plukenetione A to the antitumoral activity of Cuban propolis. Bioorg. Med. Chem., 16: 9635- 9643.
Dobrowolski, J. W., S. B. Vohora, K. Sharma, S. A. Shah, S. A. Naqvi and P. C. Dandiya (1991). Antibacterial, antifungal, antiamoebic, antiinflammatory, antipyretic studies on propolis bee products. J. Ethnopharmacol., 35: 77-82.
Eeken, J. C., I. Klink, B. L. van Veen and A. Pastink (2002). Induction of epithelial tumors in Drosophila melanogaster heterozygous for the tumor suppressor gene wts. Environ. Mol. Mutagen, 40: 277-282.
Eom, H. S., E. J. Lee, B. S. Yoon and B. S. Yoo (2010). Propolis inhibits the proliferation of human leukaemia HL-60 cells by inducing apoptosis through the mitochondrial pathway. Nat. Prod. Res., 24: 375-386.
Falcão, S. I., M. Vilas-Boas, L. M. Estevinho, C. Barros, M. R. M. Domingues and S. M. Cardoso (2010). Phenolic characterization of Northeast Portuguese propolis: usual and unusual compounds. Analytical and Bioanalytical Chemistry, 396: 887-897.
Femia, A. P., G. Caderni, C. Buzzigoli, E. Cocca, M. Salvadori and P. Dolara (2001). Effect of simple phenolic compounds on azoxymethaneinduced aberrant crypt foci in rat colon. Nutrition and Cancer, 41: 107-110.
Fly, B. (2008). A database for the Drosophila research community. Drysdale R. and The FlyBase Consortium, Methods Mol. Biol., 420: 45- 59.
Fry, F. H. and C. Jacob (2006). Sensor/ effector drug design with potential relevance to cancer. Curr. Pharm., 12: 4479-4499.
Ghisalberti, E. L. (1979). Propolis. A review. Bee World, 60: 59-84.
Hayashi, A., A. C. Gillen and J. R. Lott (2000). Effects of daily oral administration of quercetin chalcone and modified citrus pectin on implanted colon tumor growth in Balb-c mice. Alternative Medicine Review, 5: 546-552.
Hegazi, A. G. and F. K. Abd El Hady (2001). Egyptian propolis. 1. Antimicrobial activity and chemical composition of Upper Egypt propolis. Z. Naturforsch., 56: 82-88
Hegazi, A. G. and F. K. Abd El Hady (2002). Egyptian propolis: 3. Antioxidant, antimicrobial activity and chemical composition of propolis from reclaimed land. Z. Naturforsch., 57: 395-402.
Hegazi, A. G., A. M. Abdou and F. Abd Allah (2014). Egyptian Propolis 11: Its antimicrobial activity with comparison with different localities. Int. J. Curr. Microbiol. App. Sci., 3: 530-538.
Ibrahim, T., S. Demir, S. Misir, K. Kilinc, A. Mentese, Y. Aliyazicioglu and O. Deger (2015). Cytotoxic effect of Turkish propolis on liver, colon, breast, cervix and prostate cancer cell lines. Tropical Journal of Pharmaceutical Research, 14: 777- 782.
Ivanauskas, L., V. Jakstas, J. Radusiene, A. Lukosius and A. Baranauskas (2008). Evaluation of phenolic acids and phenylpropanoids in the crude drugs. Medicina, 44: 48-55.
Jin, U. H., K. H. Song, M. Motomura, I. Suzuki, Y. H. Gu, Y. J. Kang, T. C. Moon and C. H. Kim (2008). Caffeic acid phenethyl ester induces mitochondria-mediated apoptosis in human myeloid leukemia U937 cells. Mol. Cell Biochem., 310: 43-48.
Kalia, P., Kumar, R. K. and K. Harjai (2013). Phytochemical screening and antibacterial activity of different extracts of Propolis. International Journal of Pharmaceutical and Biological Research, 3: 219- 222.
Kamiya, T., H. Nishihara, H. Hara and T. Adachi (2012). Ethanol extract of Brazilian red propolis induces apoptosis in human breast cancer MCF‑ 7 cells through endoplasmic reticulum stress. J. Agric. Food Chem., 60: 11065-11070.
Kędzia, B. (2009). Chemical composition of polish propolis. Part I.: The initial period of investigations. Post. Fitoter., 1: 39-44.
Kumazawa, S., M. R. Ahn, T. Fujimoto and M. Kato (2010). Radicalscavenging activity and phenolic constituents of propolis from different regions of Argentina. Natural Product Research, 24: 804-812.
Matsuno, T. (1995). A new clerodane diterpenoid isolated from propolis. Zeitschrift fur Naturforschung, 50: 93-97.
Matsuno, T., C. Chen and P. Basnet (1997). A tumoricidal and antioxidant compound isolated from an aqueous extract of propolis. Medical Science Research, 25: 583-584.
Moreira, L., L.G. Dias, J. A. Pereira and L. Estevinho (2008). Antioxidant properties, total phenols and pollen analysis of propolis samples from Portugal. Food Chem. Toxicol., 46: 3482-3485.
Mosmann, T. (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods, 65: 55-63.
Mouse, H. A., M. Tilaoui, A. Jaafari, L. A. Mbarek, R. Aboufatima and A. Chait (2012). Evaluation of the in vitro and in vivo anticancer properties of Moroccan propolis extracts. Rev. Bras. Farmacogn., 22: 558- 567.
Nepomuceno, J. C. (2015). Using the Drosophila melanogaster to assessment carcinogenic agents through the test for detection of epithelial tumor clones (Warts). Adv. Tech. Biol. Med., 3: 149-156.
Newairy, A. A., A. F. Salama, H. M. Hussien and M. I. Yousef (2009). Aluminium induced alterations in biochemical parameters and lipid peroxidation of male rats: protective role of propolis. Food Chem. Toxicol., 47: 1093-1098
Oršolić, N. and I. Basic (2007). Cancer chemoprevention by propolis and its polyphenolic compounds in experimental animals. Recent Progress in Medicinal Plants, 17: 55- 113.
Oršolić, N., A. H. Knezevic, L. Sver, S. Terzic and Basic, I. (2004). Immunomodulatory and antimetastatic action of propolis and related polyphenolic compounds. J. Ethnopharmacol. 94: 307-315.
Oršolić, N., S. Terzić, Ž. Mihaljević, L. Šver and I. Bašić (2005). Effects of local administration of propolis and its polyphenolic compounds on tumor formation and growth. Biological and Pharmaceutical Bulletin., 28: 1928-1933.
Orsolin, P. C., R. G. Silva-Oliveira and J. C. Nepomuceno (2012). Assessment of the mutagenic, recombinagenic and carcinogenic potential of orlistat in somatic cells of Drosophila melanogaster. Food Chem. Toxicol., 50: 2598-2604.
Park, Y. K., S. M. Alencar and C. L. Aguiar (2002). Botanical origin and chemical composition of Brazilian propolis. J. Agric. Food Chem., 50: 2502-2506.
Popova, M., S. Silici, O. Kaftanoglu and V. Bankova (2005). Antibacterial activity of Turkish propolis and its qualitative and quantitative chemical composition. Phytomedicine, 12: 221-228.
Ramanauskien, K., A. M. Inknien, V. Petrikait and V. Briedis (2013). Total phenolic content and antimicrobial activity of different Lithuanian propolis solutions. Hindawi Publishing Corporation, Evidence- Based Complementary and Alternative Medicine, 2-5.
Repetto, G., A. Peso and J. L. Zurita (2008). Neutral red uptake assay for the estimation of cell viability/ cytotoxicity. Nature Protocols, 3: 1152-1135.
Salonen, A., S. Saarnio and R. Julkunen- Tiitto (2012). Phenolic compounds of propolis from the boreal coniferous zone. Journal of Agicultural Science, 56: 13-22.
Sawicka, D., H. Car, M. H. Borawska and J. Niklinski, (2012). The anticancer activity of proplis. Folia Histochemica et Cytobiologica, 50: 25-37.
Scheller, S., W. Krol, J. Swiacik, S. Owczarek, J. Gabrys and J. Shani (1989) Antitumoral property of ethanolic extract of propolis in mice-bearing Ehrlich carcinoma, as compared to bleomycin. Zeitschrift fur Naturforschung, 44c: 1063- 1065.
Sforcin, J. M. (2007). Propolis and the immune system: A Review. J. Ethnopharmacol., 113: 1-14.
Sibel, S. and K. Semiramis (2005). Chemical composition and antibacterial activity of propolis collected by three different races of honeybees in the same region. Journal of Ethnopharmacology, 99: 69-73
Siddiqui, M. A., G. Singh, M. P. Kashyap, V. K. Khanna, S. Yadav, D. Chandra and A. B. Pant (2008). Influence of cytotoxic doses of 4- hydroxynonenal on selected neurotransmitter receptors in PC-12 cells. Toxicol: In Vitro, 22: 1681- 1688.
Silici, S. and K. Semiramis (2005). Chemical composition and antibacterial activity of propolis collected by three different races of honeybees in the same region. J. Ethnopharmacol., 99: 69-73.
Umthong, S., P. Phuwapraisirisan, S. Puthong and Chanchao, C. (2011). In vitro antiproliferative activity of partially purified Trigona laeviceps propolis from Thailand on human cancer cell lines. BMC Complement Altern. Med., 11: 37.
Vatansever, H. S., K. Sorkun, S. I. D. Gurhan, F. O. Kurt, E. Turkoz, O. Gencay and B. Salih (2010). Propolis from Turkey induces apoptosis through activating caspases in human breast carcinoma cell lines. Acta Histochem., 112: 546-556.
Watabe, M., K. Hishikawa, A. Takayanagi, N. Shimizu and T. Nakaki (2004). Caffeic acid phenethyl ester induces apoptosis by inhibition of NFkappaB and activation of Fas in human breast cancer MCF-7 cells. J. Biol. Chem., 279: 6017-6026.
Watanabe, M. A. E., M. K. Amarante, B. J. Conti and J. M. Sforcin (2011). Cytotoxic constituents of propolis inducing anticancer effects: A review. J. Pharm. Pharmacol., 63: 1378-1386.
Yusuf, N., C. Irby, S. K. Katiyar and C. A. Elmets (2007). Photoprotective effects of green tea polyphenols. Photodermatol Photoimmunol Photomed, 23: 48-56.
Zizic, J. B., N. L. Vukovic, M. B. Jadranin, B. D. Andelkovic, V. V. Tesevic, M. M. Kacaniova, S. B. Sukdolak and S. D. Markovic (2013). Chemical composition, cytotoxic and antioxidative activities of ethanolic extracts of propolis on HCT-116 cell line. J. Sci. Food Agric., 93: 3001-3009.
