Evaluation of cytotoxic effect of different Iranian Chrysanthemum morifolium cultivars against MCF7 cell lines

Evaluation of cytotoxic effect of different Iranian Chrysanthemum morifolium cultivars against MCF7 cell lines
Mahboobeh Hodaei,^1,* Mandana Behbahani,² Mehdi Rahimmalek,³
1. Isfahan University of Technology
2. University of Isfahan
3. Isfahan University of Technology
Introduction: Cancer is the second leading cause of death in the world (Khakdan and Piri, 2013). Breast cancer is the most common disease in women, which can be treated with natural therapy (Cushman & Nagarathnam, 1991). Despite the availability of synthetic drugs, plants still are one of the important sources for modern drug development for anticancer activity (Kashif et al., 2014), as over 50% of anticancer drugs used in therapical trials were isolated from natural sources or are related to them . The medicinal value of plants depends on some phytochemicals (Khakdan and Piri, 2013) like terpenes, phenolics, and alkaloids that produce a physiological action on the human body (Dai & Mumper, 2010: Kashif et al., 2014). Chrysanthemum morifolium has been used as an herbal in traditional medicine applications for several years. Recently, it had been reported that Chrysanthemum flavonoids showed cytotoxic effect against human breast cancer cells (Liu, Mou, Zhou, Zhou & Shou, 2018). In Iran, despite the widespread ornamental use of these herbal plants, no scientific assessment has been reported for anticancer effect of them. considering the anti-cancer effect of this plant, this research was conducted to investigate cytotoxic and growth inhibitory activities of C. morifolium flower extract on human breast cancer cell lines (MCF7).
Methods: Sixteen cultivars of Chrysanthemum (Chrysanthemum morifolium Ramat.) including Shokoh, Atashgoon, Atash2, Sahar, Marmar, Sahand2, Dorna2, Ashna, Hour, Erica, Poya3, Bolor, Romina, Farhood, Taraneh, and Mehrnoosh2 originated from the Iranian Research Center for Ornamental Plants, Mahallat, Iran were used in this study. Dried ground flowers (2.5 g) was extracted with methanol-water (50ml, 80:20, v/v) and orbital shaker for 24 h. The extract was filtered through three layers of cheesecloth. The filtered extracts were evaporated under reduced pressure to obtain dried residue and after that a stock solution was prepared by dissolving the extract powders in DMSO to form a concentration of 100 mg/ml. Human cancer cell lines (line MCF7) were cultured in RPMI-1640 media. Three wells for each concentration were seeded. Cells were maintained in a humidified 5% CO2 incubator at 37 °C. The effect of Chrysanthmeum extracts on MCF7 cells were determined by 3-(4,5 dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay. Cell viability was calculated using the following formula: % Cells viability= (OD sample/OD control) ×100. The data represent mean ± SE from triplicate experiments. One-way analysis of variance (ANOVA) was carried out and Fisher’s (protected) least significant difference (LSD) was applied to compare means at the 95% confidence level using SAS ver. 9 software.
Results: The results demonstrated that all tested extracts exhibited different potency of cytotoxic activity against the MCF7 cell lines at five various concentrations. All cultivars showed maximum cytotoxic effect at 0.312 mg/ml concentration which the methanol extracts of cultivars “Dorna2” and “Farhood” inhibited the viability of the cancer cell lines by up to 50%. Murayama et al. (2012) reported that both leaf and flower water extracts of Chrysanthemum suppressed the proliferation of MCF7 cell lines in a dose dependent manner at concentrations >25 μg/ml. Lee et al. (2014) evaluated anticancer activity of some Korean Chrysanthemum sp. (C. boreale, C. indicum and C. morifoium). These methanolic extracts exhibited relatively potent inhibition at a concentration of 200 μg/ml against MCF7 cell lines with cell viability of 47-63%. Treatment (200 μg/ml) of C. morifoium suppressed 49% of cell viability in this study. Flavonoids have been found to possess anti-cancer activity due to their impact on the transduction in cell proliferation (Khakdan & Piri, 2013). Previous studies reported that the flavonoids from C. morifolium exhibited significant cytotoxicities against human breast, liver and colon cancer cells (Peng, Zou & Xu, 2010; Xie, Yuan, Yang, Wang & Wu, 2009). So, the observed anti-cancer effect of the extracts on the MCF7 cell lines could be attributed to the presence of these bioactive compounds.
Conclusion: The results demonstrated that different Chrysanthemum cultivars could be evaluated as an anticancer agent in different concentrations, which is useful in the cytotoxic evaluation of new herbal drugs. The present study established that the Iranian Chrysanthemum cultivars can be considered as promising sources of anti-cancer agent.
Keywords: Chrysanthemum, Breast cancer, MCF7, Toxicity, Herbal drug