Cancer Letters

Cancer Letters

Volume 236, Issue 2, 18 May 2006, Pages 282-291
Cancer Letters

In vitro antileukaemic activity of extracts from berry plant leaves against sensitive and multidrug resistant HL60 cells

https://doi.org/10.1016/j.canlet.2005.05.018Get rights and content

Abstract

The aim of the present study was to determine in vitro antileukaemic activity of extracts obtained from selected berry plant leaves (Fragaria x ananassa Duch. cv Elsanta, raspberry Rubus ideus L. cv Polana and blueberry Vaccinium corymbosum L. cv Bluecrop) against promyelocytic HL60 cell line and its multidrug resistant sublines exhibiting two different MDR phenotypes: HL60/VINC (overexpressing P-glycoprotein) and HL60/DOX (overexpressing MRP1 protein). It was found that the blueberry extract was the most efficient against sensitive HL60 cell line (about 2-fold more active than strawberry and raspberry extracts) but presented much lower activity towards resistant cells. In contrast, strawberry and raspberry extracts exhibited the high cytotoxic activity against sensitive leukaemia HL60 cell line as well as its MDR sublines. The values of resistance factor (RF) found for these extracts were very low lying in the range 0.32÷2.0.

Introduction

The food of plant origin, especially fruits and vegetables draw increased attention because of its potential benefits to human health. Epidemiological studies showed that there is a significant positive correlation between intake of fruits and vegetables and reduced rate of heart diseases, cancer mortality and degenerative diseases [1], [2], [3], [4]. Phytochemicals, especially phenolics, in fruits and vegetables are suggested to be the major bioactive compounds for the health benefits because they act as antioxidants and scavenge free radicals [5].

Recently, the attempts of many researchers are focused on the identification of phytochemicals able to inhibit cancer cell growth. There is an increasing number of evidence indicating in vitro antiproliferative activity of fruit extracts against various cancer cell lines. Hibasami et al. [6] observed that the growth of human stomach cancer KATO III cells was inhibited by genistein of soybean koji. It was also found that green tea catechins: (−)-epicatechin, (−)-epigallocatechin and (−)-epigallocatechin gallate inhibited the growth of HCT 116 colorectal and HepG2 hepatocellular carcinoma cells [7]. There is also a recent study showing that theaflavin digallate (black tea), epigallocatechin gallate (green tea) and oolong tea polyphenol trimer caused the growth inhibition and induction of apoptosis in human colon cancer COLO 205 cells [8]. Saleem et al. [9] observed antiproliferative effect of fruit extract of Terminalia chebula on human prostate cancer PC-3, human MCF-7 and mouse S115 breast cancer and human osteosarcoma HOS-1 cell lines. It is also proposed that some phenolics namely flavonoids: catechins, quercetin, kaempferol, anthocyanins and phenolic acids: derivatives of hydroxybenzoic and hydroxycinnamic acids, especially ellagic acid could exibit a high antiproliferative activity against cancer cells [10], [11], [12], [13]. These phytochemicals are abundantly occurring in fruits and vegetables. Published data demonstrated that the ability of phenolic compounds to inhibit the growth of leukaemic and carcinoma cells could be related to the G1 or G2/M phase arrest as well as the induction of apoptosis through cytochrome c release and activation of caspase 9 and caspase 3 [14], [15], [16], [17].

However, one of the most important problems in cancer chemotherapy is the occurrence of multidrug resistance (MDR) associated with the presence of membrane transporters (e.g. P-glycoprotein, MRP1), belonging to the ATP-binding cassette protein family [18], [19], [20], [21]. Some reports presented the properties of plant compounds (e.g. genistein, green tea polyphenols, compounds present in bitter melon Momocardia charantia extracts) as reversal agents able to restore the activity of antitumour agents, especially doxorubicin and daunorubicin against multidrug resistant tumour cells overexpressing P-glycoprotein [22], [23], [24], [25]. In contrast, it should be noted that although there are numerous data demonstrating the ability of phytochemicals to inhibit themselves the growth of various cancer cells, up to now very little is known about their antiproliferative activity towards multidrug resistant tumour cells. Thus, the search for substances better protecting against cancer is ongoing all over the world. Recent study showed that the pentacyclic triterpenes isolated from Chrysobalanaceae species inhibited the growth and induced apoptosis of erythroleukaemia K562 cell line and its multidrug resistant subline K562/DOX overexpressing P-glycoprotein [26]. Apart from edible fruits and vegetables, other plant materials as the sources of biologically active substances, especially rich in ellagic acid are studied: leaves and other parts of pomegranate [27], fruits of Terminalia sp. or leaves of Alangium javanicum, branches of Anisophyllea apetala, flowers, leaves and twigs of Crypteronia paniculata, roots of Mouririi sp., stem bark and stem wood of Schlotzia parviflora [28]. Also non-edible parts of strawberry: green fruits, achenes and leaves were found richer in ellagic acid than ripe fruits [29], [30]. Likewise, it was evidenced that young leaves of strawberry, raspberry and blackberry had higher total phenolic content than the respective fruit tissues and old leaves [31].

The aim of the present study was to determine in vitro antileukaemic activity of extracts obtained from selected berry plant leaves against promyelocytic HL60 cell line and its multidrug resistant sublines exhibiting two different MDR phenotypes: HL60/VINC (overexpressing P-glycoprotein) and HL60/DOX (overexpressing MRP1 protein).

Section snippets

Chemicals

HCl, NaOH and methanol were of analytical grade and purchased from POCh Gliwice, Poland. HPLC chemicals were of HPLC purity and purchased from Sigma–Aldrich (USA).

Cell culture

HL60 human promyelocytic leukemia (Division of Biology, Kansas State University, Manhattan, KS 66506, USA) and their resistant sublines: HL-60/VINC (overexpressing P-glycoprotein) [32] and HL-60/DOX (overexpressing MRP1) [33], [34] were cultured. The cells were grown in RPMI 1640 (Gibco Limited) medium supplemented with 2 mM glutamine

Composition of strawberry, raspberry and blueberry leaf extracts

The dry weight of strawberry, raspberry and blueberry leaves used for extract preparation is shown in Table 1. The highest dry weight content in fresh material was observed in blueberry leaves (39.51±1.97 g/100 g) and the lowest in strawberry leaves (32.85±1.64 g/100 g). In contrast, the values of dry weight content of the leaves after room temperature air-drying were very similar for the three berry plant species studied (about 92–93 g/100 g).

The representative chromatograms of phenolic compounds

Discussion

Multidrug resistance (MDR) constitutes the major problem in cancer therapy. Tumour cells become resistant to a wide array of chemotherapeutic agents (e.g. anthracyclines, vinca alkaloids, podophylotoxins, colchicine), structurally diverse and having different mechanisms of action [35]. Thus, many efforts are focused on the search of agents conserving the antitumour activity against multidrug resistant cells [36], [37], [38], [39]. Alternatively, the identification of effective chemosensitizers

Acknowledgements

This work was supported by the grant BW/HK/11/2004 of Agricultural Academy of Szczecin, Poland. The authors wish to thank Katarzyna Cholewińska, MSc for supplying strawberry and raspberry leaves as well as Dorota Zaborowska, MSc and Adam Zaborowski, MSc for supplying blueberry leaves for this study.

References (52)

  • A. Zhu et al.

    Study of tea polyphenol as a reversal agent for carcinoma cell lines' multidrug resistance (study of TP as a MDR reversal agent)

    Nucl. Med. Biol.

    (2001)
  • J. Fernandes et al.

    Pentacyclic triterpenes from Chrysobalanaceae species: cytotoxicity on multidrug resistant and sensitive leukemia cell lines

    Cancer Lett.

    (2003)
  • F. Lei et al.

    Pharmacokinetic study of ellagic acid in rat after oral administration of pomegranate leaf extract

    J. Chromatogr. B.

    (2003)
  • Y. Xu et al.

    DNA damaging activity of ellagic acid derivatives

    Bioorg. Med. Chem.

    (2003)
  • T. McGrath et al.

    Mechanisms of multidrug resistance in HL60 cells. Analysis of resistance associated membrane proteins and levels of mdr gene expression

    Biochem. Pharmacol.

    (1989)
  • J.L. Donovan et al.

    Catechin is metabolized by both the small intestine and liver of rats

    J. Nutr.

    (2001)
  • S.A. Aherne et al.

    Dietary flavonols: chemistry, food content, and metabolism

    Nutrition

    (2002)
  • J.M. Gee et al.

    Intestinal transport of quercetin glycosides in rats involves both deglycosylation and interaction with the hexose transport pathway

    J. Nutr.

    (2000)
  • F. Lei et al.

    Pharmacokinetic study of ellagic acid in rat after oral administration of pomegranate leaf extract

    J. Chromatogr. B

    (2003)
  • S.U. Mertens-Talcott et al.

    Ellagic acid and quercetin interact synergistically with resveratrol in the induction of apoptosis and cause transient cell cycle arrest in human leukemia cells

    Cancer Lett.

    (2005)
  • S.U. Mertens-Talcott et al.

    Low concentrations of quercetin and ellagic acid synergistically influence proliferation, cytotoxicity and apoptosis in MOLT-4 human leukemia cells

    J. Nutr.

    (2003)
  • P.C.H. Hollman et al.

    Analysis and health effects of flavonoids

    Food Chem.

    (1996)
  • Y.S. Velioglu et al.

    Antioxidant activity and total phenolics content in selected fruits, vegetables, and grain products

    J. Agric. Food Chem.

    (1998)
  • C. Kaur et al.

    Antioxidants in fruits and vegetables—the millenium's health

    Int. J. Food Sci. Technol.

    (2001)
  • H. Hibasami et al.

    Human stomach cancer cells undergo apoptosis by genistein, but not other genistein- and daidzein-related compounds

    J. Herbs, Spices Med. Plants

    (2003)
  • H. Hibasami et al.

    Human colon cancer cells undergo apoptosis by theaflavin digallate, epigallocatechin gallate, and oolong tea polyphenol extract

    J. Herbs, Spices Med. Plants

    (2003)
  • Cited by (0)

    View full text