期刊论文详细信息
BMC Complementary and Alternative Medicine
Propolis changes the anticancer activity of temozolomide in U87MG human glioblastoma cell line
Halina Car1  Diana Sawicka2  Sylwia K Naliwajko3  Anna Fiedorowicz1  Maria H Borawska3  Renata Markiewicz-Żukowska3 
[1] The Department of Experimental Pharmacology, Medical University of Bialystok, Szpitalna 37, 15-295, Bialystok, Poland;The Center of Experimental Medicine, Medical University of Bialystok, Marii Sklodowskiej-Curie 24A, 15-276, Bialystok, Poland;The Department of Bromatology, Medical University of Bialystok, Mickiewicza 2D, 15-222, Bialystok, Poland
关键词: NF-κB;    DNA synthesis;    Cell viability;    Temozolomide;    Glioblastoma;    Propolis;   
Others  :  1230292
DOI  :  10.1186/1472-6882-13-50
 received in 2012-07-04, accepted in 2013-02-25,  发布年份 2013
【 摘 要 】

Background

Propolis is a honey bee product which contains many active compounds, such as CAPE or chrysin, and has many beneficial activities. Recently, its anti-tumor properties have been discussed. We have tested whether the ethanolic extract of propolis (EEP) interferes with temozolomide (TMZ) to inhibit U87MG cell line growth.

Methods

The U87MG glioblastoma cell line was exposed to TMZ (10-100 μM), EEP (10-100 μg/ml) or a mixture of TMZ and EEP during 24, 48 or 72 hours. The cell division was examined by the H3-thymidine incorporation, while the western blot method was used for detection of p65 subunit of NF-κB and ELISA test to measure the concentration of its p50 subunit in the nucleus.

Results

We have found that both, TMZ and EEP administrated alone, had a dose- and time-dependent inhibitory effect on the U87MG cell line growth, which was manifested by gradual reduction of cell viability and alterations in proliferation rate. The anti-tumor effect of TMZ (20 μM) was enhanced by EEP, which was especially well observed after a short time of exposition, where simultaneous usage of TMZ and EEP resulted in a higher degree of growth inhibition than each biological factor used separately. In addition, cells treated with TMZ presented no changes in NF-κB activity in prolonged time of treatment and EEP only slightly reduced the nuclear translocation of this transcription factor. In turn, the combined incubation with TMZ and EEP led to an approximately double reduction of NF-κB nuclear localization.

Conclusions

We conclude that EEP presents cytotoxic properties and may cooperate with TMZ synergistically enhancing its growth inhibiting activity against glioblastoma U87MG cell line. This phenomenon may be at least partially mediated by a reduced activity of NF-κB.

【 授权许可】

   
2013 Markiewicz-Zukowksa et al; licensee BioMed Central Ltd.

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【 参考文献 】
  • [1]Brada M, Hoang-Xuan K, Rampling R, Dietrich PY, Dirix LY, Macdonald D, Heimans JJ, Zonnenberg BA, Bravo-Marques JM, Henriksson R, Stupp R, Yue N, Bruner J, Dugan M, Rao S, Zaknoen S: Multicenter phase II trial of temozolomide in patients with glioblastoma multiforme at first relapse. Ann Oncol 2001, 12:259-266.
  • [2]Marchesi F, Turriziani M, Tortorelli G, Avvisati G, Torino F, De Vecchis L: Triazene compounds: mechanism of action and related DNA repair systems. Pharmacol Res 2007, 56:275-287.
  • [3]Aoki T, Mizutani T, Ishikawa M, Sugiyama K, Hashimoto N: A first feasibility study of temozolomide for Japanese patients with recurrent anaplastic astrocytoma and glioblastoma multiforme. Int J Clin Oncol 2003, 8:301-304.
  • [4]Kim JT, Kim JS, Ko KW, Kong DS, Kang CM, Kim MH, Son MJ, Song HS, Shin HJ, Lee DS, Eoh W, Nam DH: Metronomic treatment of temozolomide inhibits tumor cell growth through reduction of angiogenesis and augmentation of apoptosis in orthotopic models of gliomas. Oncol Rep 2006, 16:33-39.
  • [5]Uzzaman M, Keller G, Germano IM: Enhanced proapoptotic effects of tumor necrosis factor-related apoptosis-inducing ligand on temozolomide-resistant glioma cells. J Neurosurg 2007, 106:646-651.
  • [6]Maciejewicz W, Daniewski M, Bal K, Markowski W: GC-MS identification of the flavonoid aglycones isolated from propolis. Chromatogr 2001, 53:343-346.
  • [7]Vandar-Unlu G, Silici S, Unlu M: Composition and in vitro antimicrobial activity of Populus buds and poplar-type propolis. World J Microbiol Biotechnol 2008, 24:1011-1017.
  • [8]Marcucci MC: Propolis: chemical composition, biological properties and therapeutic activity. Apidologie 1995, 26:88-99.
  • [9]Kujumgiev A, Tsvetkova I, Serkedjieva Y, Bankova V, Christov R, Popov S: Antibacterial, antifungal and antiviral activity of propolis of different geographic origin. J Ethnopharmacol 1999, 64:235-240.
  • [10]Silici S, Unlu M, Vardar-Unlu G: Antibacterial activity and phytochemical evidence for the plantorigin of Turkish propolis from different regions. World J Microbiol Biotechnol 2007, 23:1797-1803.
  • [11]Amoros M, Sauvager F, Girre L, Cormier M: In vitro antiviral activity of propolis. Apidologie 1992, 23:231-240.
  • [12]EFSA: Panel on Dietetic Products, Nutrition and Allergies (NDA). EFSA Jurnal 2010, 180:1-16.
  • [13]Daugsch A, Moraes CS, Fort P, Park YK: Brazilian red propolis-chemical composition and botanical origin. Evid Based Complement. Alternat Med 2008, 5:435-441.
  • [14]Chen CN, Hsiao CJ, Lee SS, Guh JH, Chiang PC, Huang CC, Huang WJ: Chemical modification and anticancer effect of prenylated flavanones from Taiwanese propolis. Nat Prod Res 2012, 26:116-124.
  • [15]Chen YJ, Shiao MS, Hsu ML, Tsai TH, Wang SY: Effect of caffeic acid phenethyl ester, an antioxidant from propolis, on inducing apoptosis in human leukemic HL-60 cells. J Agric Food Chem 2001, 49:5615-5619.
  • [16]Nomura M, Kaji A, Ma W, Miyamoto K, Dong Z: Suppression of cell transformation and induction of apoptosis by caffeic acid phenethyl ester. Mol Carcinog 2001, 31:83-89.
  • [17]Watabe M, Hishikawa K, Takayanagi A, Shimizu N, Nakaki T: Caffeic acid phenethyl ester induces apoptosis by inhibition of NFkappaB and activation of Fas in human breast cancer MCF-7 cells. J Biol Chem 2004, 279:6017-6026.
  • [18]Jin UH, Song KH, Motomura M, Suzuki I, Gu YH, Kang YJ, Moon TC, Kim CH: Caffeic acid phenethyl ester induces mitochondria-mediated apoptosis in human myeloid leukemia U937 cells. Mol Cell Biochem 2008, 310:43-48.
  • [19]Chen MJ, Chang WH, Lin CC, Liu CY, Wang TE, Chu CH, Shih SC, Chen YJ: Caffeic acid phenethyl ester induces apoptosis of human pancreatic cancer cells involving caspase and mitochondrial dysfunction. Pancreatology 2008, 8:566-576.
  • [20]Borges KS, Brassesco MS, Scrideli CA, Soares AE, Tone LG: Antiproliferative effects of Tubi-bee propolis in glioblastoma cell lines. Genet Mol Biol 2011, 34:310-314.
  • [21]Carmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell JB: Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res 1987, 47:936-942.
  • [22]Portnow J, Badie B, Chen M, Liu A, Blanchard S, Synold TW: The neuropharmacokinetics of temozolomide in patients with resectable brain tumors: potential implications for the current approach to chemoradiation. Clin Cancer Res 2009, 15:7092-7098.
  • [23]Rosso L, Brock CS, Gallo JM, Saleem A, Price PM, Turkheimer FE, Aboagye EO: A new model for prediction of drug distribution in tumor and normal tissues: pharmacokinetics of temozolomide in glioma patients. Cancer Res 2009, 69:120-127.
  • [24]Golden EB, Lam PY, Kardosh A, Gaffney KJ, Cadenas E, Louie SG, Petasis NA, Chen TC, Schönthal AH: Green tea polyphenols block the anticancer effects of bortezomib and other boronic acid-based proteasome inhibitors. Blood 2009, 113:5927-5937.
  • [25]Cardoso SM, Ribeiro M, Ferreira IL, Rego AC: Northeast Portuguese propolis protects against staurosporine and hydrogen peroxide-induced neurotoxicity in primary cortical neurons. Food Chem Toxicol 2011, 49:2862-2868.
  • [26]Shimazawa M, Chikamatsu S, Morimoto N, Mishima S, Nagai H, Hara H: Neuroprotection by Brazilian Green Propolis against in vitro and in vivo ischemic neuronal damage. Evid Based Complement Alternat Med 2005, 2:201-207.
  • [27]Tentori L, Graziani G: Pharmacological strategies to increase the antitumor activity of methylating agents. Curr Med Chem 2002, 9:1285-1301.
  • [28]Blough MD, Beauchamp DC, Westgate MR, Kelly JJ, Cairncross JG: Effect of aberrant p53 function on temozolomide sensitivity of glioma cell lines and brain tumor initiating cells from glioblastoma. J Neurooncol 2011, 102:1-7.
  • [29]Watanabe MA, Amarante MK, Conti BJ, Sforcin JM: Cytotoxic constituents of propolis inducing anticancer effects: a review. J Pharm Pharmacol 2011, 63:1378-1386.
  • [30]Aso K, Kanno S, Tadano T, Satoh S, Ishikawa M: Inhibitory effect of propolis on the growth of human leukemia U937. Biol Pharm Bull 2004, 27:727-730.
  • [31]Ishihara M, Naoi K, Hashita M, Itoh Y, Suzui M: Growth inhibitory activity of ethanol extracts of Chinese and Brazilian propolis in four human colon carcinoma cell lines. Oncol Rep 2009, 22:349-354.
  • [32]Awale S, Li F, Onozuka H, Esumi H, Tezuka Y, Kadota S: Constituents of Brazilian red propolis and their preferential cytotoxic activity against human pancreatic PANC-1 cancer cell line in nutrient-deprived condition. Bioorg Med Chem 2008, 16:181-189.
  • [33]Josipovic P, Orsolic N: Cytotoxicity of polyphenolic/flavonoid compounds in a leukemia cell culture. Arh Hig Rada Toksikol 2008, 59:299-308.
  • [34]Orsolic N, Sver L, Terzić S, Basić I: Peroral application of water-soluble derivative of propolis (WSDP) and its related polyphenolic compounds and their influence on immunological and antitumour activity. Vet Res Commun 2005, 29:575-593.
  • [35]Zhang R, Banik NL, Ray SK: Combination of all-trans retinoic acid and interferon-gamma suppressed PI3K/Akt survival pathway in glioblastoma T98G cells whereas NF-kappaB survival signaling in glioblastoma U87MG cells for induction of apoptosis. Neurochem Res 2007, 32:2194-2202.
  • [36]Fukushima T, Kawaguchi M, Yorita K, Tanaka H, Takeshima H, Umezawa K, Kataoka H: Antitumor effect of dehydroxymethylepoxyquinomicin, a small molecule inhibitor of nuclear factor-κB, on glioblastoma. Neuro Oncol 2012, 14:19-28.
  • [37]Ha SK, Moon E, Kim SY: Chrysin suppresses LPS-stimulated proinflammatory responses by blocking NF-κB and JNK activations in microglia cells. Neurosci Lett 2010, 485:143-147.
  • [38]Vallabhapurapu S, Karin M: Regulation and function of NF-kappaB transcription factors in the immune system. Annu Rev Immunol 2009, 27:693-733.
  • [39]Chaturvedi MM, Sung B, Yadav VR, Kannappan R, Aggarwal BB: NF-κB addiction and its role in cancer: ‘one size does not fit all’. Oncogene 2011, 30:1615-1630.
  • [40]Yang J, Splittgerber R, Yull FE, Kantrow S, Ayers GD, Karin M, Richmond A: Conditional ablation of Ikkb inhibits melanoma tumor development in mice. J Clin Invest 2010, 120:2563-2574.
  • [41]Cuenin S, Tinel A, Janssens S, Tschopp J: p53-induced protein with a death domain (PIDD) isoforms differentially activate nuclear factor-kappaB and caspase-2 in response to genotoxic stress. Oncogene 2008, 27:387-396.
  • [42]Kim EL, Yoshizato K, Kluwe L, Meissner H, Warnecke G, Zapf S, Westphal M, Deppert W, Giese A: Comparative assessment of the functional p53 status in glioma cells. Anticancer Res 2005, 25:213-224.
  • [43]Amin AR, Khuri FR, Chen ZG, Shin DM: Synergistic growth inhibition of squamous cell carcinoma of the head and neck by erlotinib and epigallocatechin-3-gallate: the role of p53-dependent inhibition of nuclear factor-kappaB. Cancer Prev Res (Phila) 2009, 2:538-545.
  • [44]Hirose Y, Berger MS, Pieper RO: p53 effects both the duration of G2/M arrest and the fate of temozolomide-treated human glioblastoma cells. Cancer Res 2001, 61:1957-1963.
  • [45]Watt HL, Rachid Z, Jean-Claude BJ: Receptor activation and inhibition in cellular response to chemotherapeutic combinational mimicries: the concept of divergent targeting. J Neuro-Oncol 2010, 100:345-361.
  • [46]Lu W, Zhou X, Hong B, Liu J, Yue Z: Suppression of invasion in human U87 glioma cells by adenovirus-mediated co-transfer of TIMP-2 and PTEN gene. Cancer Lett 2004, 214:205-213.
  • [47]Maehama T, Dixon JE: PTEN: a tumour suppressor that functions as a phospholipid phosphatase. Trends Cell Biol 1999, 9:125-128.
  • [48]Zhang X, Chen T, Zhang J, Mao Q, Li S, Xiong W, Qiu Y, Xie Q, Ge J: Notch1 promotes glioma cell migration and invasion by stimulating β-catenin and NF-κB signaling via AKT activation. Cancer Sci 2012, 103:181-190.
  • [49]Chen L, Han L, Shi Z, Zhang K, Liu Y, Zheng Y, Jiang T, Pu P, Jiang C, Kang C: LY294002 enhances cytotoxicity of temozolomide in glioma by down-regulation of the PI3K/Akt pathway. Mol Med Report 2012, 5:575-579.
  • [50]Wang Y, Chen L, Bao Z, Li S, You G, Yan W, Shi Z, Liu Y, Yang P, Zhang W, Han L, Kang C, Jiang T: Inhibition of STAT3 reverses alkylator resistance through modulation of the AKT and β-catenin signaling pathways. Oncol Rep 2011, 26:1173-1180.
  • [51]Kim YH, Lee YJ: TRAIL apoptosis is enhanced by quercetin through Akt dephosphorylation. J Cell Biochem 2007, 100:998-1009.
  • [52]Wang LC, Chu KH, Liang YC, Lin YL, Chiang BL: Caffeic acid phenethyl ester inhibits nuclear factor-kappaB and protein kinase B signalling pathways and induces caspase-3 expression in primary human CD4+ T cells. Clin Exp Immunol 2010, 160:223-232.
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