Cytotoxic response of breast cancer cell lines, MCF 7 and T 47 D to triphala and its modification by antioxidants
Introduction
Breast cancer is the second most common cancer in Indian women. Every year, over 80,000 new cases of breast cancer are diagnosed in India and is one of the leading causes of death in women. This implies that there is a continuing need for development of new anticancer drugs, drug combinations and treatment modalities to effectively treat breast cancer patients. Natural products, especially plant-based products have frequently been examined as anti cancer agents [1]. There exists high hope for effective treatment of different cancers by systematic screening of a variety of natural products. Ayurveda, the ancient Indian science of health, is based on natural products including several phytochemicals for treatment of a variety of diseases. Reports have appeared showing that plants extracts displayed anti-tumor/anti-cancer/anti-proliferative effects on cultured human tumor cell lines [2], [3], [4], [5], [6], [7]. Some of the recent reports, concerned with, purified compounds from herbal extracts with their possible mechanisms of anti-cancer effects [8], [9], [10], [11], [12], [13]. Polyherbal formulations from Chinese and Indian medicinal systems have long been advocated for anticancer properties [14], [15], [16]. One such formulation from Ayurveda, Triphala (TPL), extracted in acetone has been recently reported for its anticancer property [17].
TPL is an herbal drug consisting of the dried and powdered fruits of three plants, Terminalia chebula, Emblica officinalis and Terminalia bellerica in equal proportions. It is an important medicine of the ‘rasayana’ group and is believed to promote health, immunity and longevity [18]. This formulation, rich in antioxidants, is frequently used in Ayurvedic medicine to treat many diseases such as anemia, jaundice, constipation, asthma, fever and chronic ulcers.
We have recently shown that TPL was significantly more toxic to tumor cells in vitro as well as in vivo as compared to normal cells [19]. In addition, it has been shown that TPL induced apoptosis in tumor cells in vitro. This study describes the cytotoxic response of two breast cancer cell lines, MCF 7 (p53+ve) and T 47 D (p53−ve) to TPL as evaluated by MTT and clonogenic assays. It has further been shown that pifithrin, a known inhibitor of p53, markedly removed the TPL induced cytotoxic effect on cells. Antioxidants, such as, GSH and NAC were found to restore the drug induced cellular toxicity. Our results have shown differential sensitivity of MCF 7 and T 47 D cells to TPL in terms of proliferation and clonogenicity. Over all, these results may have implications in developing protocols for effective treatment of cancer in clinic based on the p53 and redox status of tumor cells.
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Cell culture and harvesting
Dulbecco's Modified Eagle's Medium (DMEM) was purchased from Gibco Co., USA and fetal bovine serum (FBS) was obtained from Biomedia, France. Pifithrin-α, Glutathione and N-Acetyl-l-Cysteine were purchased from Sigma, USA. MCF 7 and T 47 D cells were purchased from National Centre for Cell Science, Pune, India and they were cultured in DMEM supplemented with FCS (10%) and antibiotics (streptomycin 200 μg/ml and penicillin 100 units /ml) at 37 °C in 5% CO2 environment. Cells were harvested from 80
Effect of TPL on proliferation of MCF 7 and T 47 D
MCF 7 and T 47 D cells were treated with increasing concentration of TPL (0–50 μg/ml) and their loss of viability was assessed by the MTT assay (Fig. 1). It was found that rapid loss of cell proliferation occurred with the increasing concentration of TPL. For each concentration of TPL treatment, cells were incubated for different time periods of 24, 48, 72 and 120 h before evaluation of their proliferation. It can be seen that inhibition of cell proliferation induced by the drug was dependant on
Discussion
Chemotherapeutic drugs are known to induce cytotoxicity in tumor cells through diverse mechanisms, in which signaling events play an important role depending upon the cell type and stimulus. This study was aimed to investigate the effects of TPL on two breast cancer cell lines, MCF 7 and T47 D, which are known to differ in their p53 status. Results have shown that TPL remarkably reduced proliferation of wild-type MCF 7 cells (p53+ve) in a dose and time dependant manner (Fig. 1(A)). The effect
Acknowledgements
Sandhya Thulasidas would like to acknowledge the support of DAE fellowship for conducting her research. The authors would like to acknowledge Ajanta pharmaceuticals and Dr K.I. Priyadarshini for providing the triphala extract. Acknowledgements are also due to Dr B.N. Pandey and Dr K.M. Lathika for critical review of the manuscript.
References (39)
- et al.
Ripe fruits of Solanum nigrum L. inhibits cell growth and induces apoptosis in MCF 7 cells
Food Chem. Toxicol.
(2003) - et al.
Pharmacological effects of garlic extract
Trends Pharmacol. Sci.
(2003) - et al.
Paeoniae radix, a Chinese herbal extract, inhibit hepatoma cells growth by inducing apoptosis in a p53 independent pathway
Life Sci.
(2002) - et al.
Methanolic extract of Pereskia bleo (Kunth) DC, (Cactaceae) induces apoptosis in breast carcinoma, T47-D cell line
J. Ethnopharmacol.
(2005) - et al.
Apoptosis induced by tea polyphenols in HL-60 cells
Cancer Lett.
(1997) - et al.
The in vitro cytotoxic and apoptotic activity of Triphala—an Indian herbal drug
J. Ethnopharmacol.
(2005) - et al.
The evaluation of the radioprotective effect of Triphala (an ayurvedic rejuvenating drug) in the mice exposed to radiation
Phytomedicine
(2002) - et al.
Pifithrin-alpha suppresses p53 and protects cochlear and vestibular hair cells from cisplatin-induced apoptosis
Neuroscience
(2003) - et al.
Cytotoxic activity of hydrolyzable tannins against human oral tumor cell lines—a possible mechanism
Phytomedicine
(2000) - et al.
Reactive oxygen species and intracellular Ca2+, common signals for apoptosis induced by gallic acid
Biochem. Pharmacol.
(1998)
Prooxidant property of green tea polyphenols epicatechin and epigallocatechin-3-gallate: implications for anticancer properties
Toxicol. In Vitro
Influence of p53 and p21(WAF1) expression on sensitivity of cancer cells to cladribine
Biochem. Pharmacol.
Redox signalling and transition metals in the control of the p53 pathway
Biochem. Pharmacol.
The molecular epidemiology of p53 gene mutations in human breast cancer
Trends Genet.
Apoptosis vs. nonapoptotic cytotoxicity induced by hydrogen peroxide
Free Radic. Biol. Med.
p53 dependent apoptosis in glioma cell lines in response to hydrogen peroxide induced oxidative stress
Int. J. Biochem. Cell Biol.
Advances in cancer therapy with plant based natural products
Curr. Med. Chem.
Antitumor activity of balsam fir oil: production of reactive oxygen species induced by alpha-humulene as possible mechanism of action
Planta Med.
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Study of the safety of oral Triphala aqueous extract on healthy volunteers
2020, Journal of Integrative MedicineCitation Excerpt :The major active constituents of Triphala are gallic acid, tannin, ellagic acid and ascorbic acid [5,6]. Previous scientific studies of Triphala have reported that it possesses antioxidant [7–10], antihypercholesteremic [11,12], antidiabetic [13,14], anti-obesity [15,16], immunomodulation [2,17,18], antimicrobial [19–27], anti-inflammatory [28–30] and anticancer properties [31–38]. Other studies of the Triphala extract have shown that it did not produce any signs of toxicity at doses of 1750 and 2000 mg/kg during acute toxicity tests [39,40].
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