Effects of propolis on cell growth and gene expression in HL-60 cells

https://doi.org/10.1016/j.jep.2005.02.005Get rights and content

Abstract

Brazilian propolis obtained from honeybee hives was extracted with water or ethanol. Cell growth-inhibitory activities of these propolis extracts were found in HL-60 human myeloid leukemia cells. The extracts-induced apoptosis in the cells, which was characterized by morphological and nucleosomal DNA fragmentation analysis. The apoptosis was mainly attributed to the induction of granulocytic differentiation, which was evaluated by nitro blue tetrazolium (NBT) reducing assays and cytofluorometric analysis for the expression of cell surface marker CD11b. DNA microarray analysis was performed to examine the gene expression profiles in the propolis-treated HL-60 cells accompanied with granulocytic differentiation, which were compared with those in all-trans retinoic acid-treated cells. Several genes were up- or down-regulated. Two genes encoding S100 calcium binding protein A9 and ferritin, heavy polypeptide 1 were up-regulated, which were also confirmed by semi-quantitative reverse transcriptase-PCR (RT-PCR). Propolis-induced growth inhibition in HL-60 cells was, at least in part, due to differentiation with gene expression profiles, which are similar to those induced by all-trans retinoic acid.

Introduction

Propolis is a resinous hive product collected from plant materials by honeybees and is considered to be a protective barrier against the bee enemies. Propolis has been used as a folk medicine and has been reported to possess therapeutic or preventive effect in inflammation, heart disease, diabetes mellitus, hepatotoxicity, and cancer (Burdock, 1998, Banskota et al., 2001). Propolis contains more than 300 different constituents including benzoic acids, flavonoids and cinnamic acid derivatives (Marcucci, 1995, Bankova et al., 2000). Although some of the observed physiological activities of propolis could be attributed to the identified chemicals, they have rarely been used independently or individually, but propolis as a complex mixture has been commonly used for health food and beverages.

In the course of studies on anticancer activity, we examined the cytotoxic effect of propolis in HL-60 human myeloid leukemia cells. HL-60 cells are known to induce differentiation into granulocytes by all-trans retinoic acid (ATRA) (Collins, 1987), which has been used for clinical therapy for acute promyelocytic leukemia. Differentiation therapy focuses on the development and use of materials designed to selectively cause terminal differentiation, leading to the eventual elimination of tumorigenic cells and rebalance of normal cellular homeostasis (Beere and Hickman, 1993). Thus, understanding the mechanisms underlying the cytotoxic activity by propolis provides a beneficial clue for the development of new drug and functional food candidates. In this study, we evaluated potency of the propolis extracts for growth suppression, which was in part due to differentiation leading to apoptosis in HL-60 cells. In addition, DNA microarray analysis was performed to examine the gene expression profiles in the propolis-treated HL-60 cells accompanied with granulocytic differentiation.

Section snippets

Materials

Brazilian propolis was collected in Minas Gerais state, where Baccharis dracunculifolia DC. is the main botanical source of the propolis (Kato et al., 2000, Midorikawa et al., 2001). Drupanin [(E)-3-prenyl-4-hydroxycinnnamic acid] and baccharin [(E)-3-prenyl-4-(2,3-dihydrocinnamoyloxy) cinnamic acid] were purified according to the method described previously (Akao et al., 2003). Artepillin C (3,5-diprenyl-4-hydroxycinnamic acid) was purchased from Wako Pure Chemical Industries (Osaka, Japan);

Effects of WEP and EEP on HL-60 cell growth and differentiation

The composition of propolis primarily depends upon the vegetation of the area from where it was collected and secondarily upon the methods for its extraction (Marcucci, 1995, Kumazawa et al., 2004). Biological activities of propolis could be related to its chemical composition (Banskota et al., 2001). Since Baccharis dracunculifolia is an important source of propolis collected from Minas Gerais (Kato et al., 2000, Midorikawa et al., 2001), which is of good quality to provide experimental

References (18)

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