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Antioxidant activity of Coriandrum sativum and protection against DNA damage and cancer cell migration

Tang, Esther LH, Rajarajeswaran, Jayakumar, Fung, Shin Yee, Kanthimathi, MS·2013·BMC Complementary and Alternative Medicine
DOI 10.1186/1472-6882-13-347PMC PMC4028854
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Abstract

Background: Coriandrum sativum is a popular culinary and medicinal herb of the Apiaceae family. Health promoting properties of this herb have been reported in pharmacognostical, phytochemical and pharmacological studies.However, studies on C. sativum have always focused on the aerial parts of the herb and scientific investigation on the root is limited. The aim of this research was to investigate the antioxidant and anticancer activities of C. sativum root, leaf and stem, including its effect on cancer cell migration, and its protection against DNA damage, with special focus on the roots. Methods: Powdered roots, leaves and stems of C. sativum were extracted through sequential extraction using hexane, dichloromethane, ethyl acetate, methanol and water. Total phenolic content, FRAP and DPPH radical scavenging activities were measured. Anti-proliferative activitiy on the breast cancer cell line, MCF-7, was assayed using the MTT assay. Activities of the antioxidant enzymes, catalase, superoxide dismutase, glutathione peroxidase, and of the caspases-3, -8 and -9 were assayed on treatment with the extract. Cell cycle progression was analysed using flow cytometry. The scratch motility assay was used to assess inhibition of MCF-7 cell migration. DNA damage in 3 T3-L1 fibroblasts was evaluated by the comet assay. The components in the extract were identified by HPLC and GC-MS. Results: The ethyl acetate extract of C. sativum roots showed the highest antiproliferative activity on MCF-7 cells (IC 50 = 200.0 ± 2.6 μg/mL) and had the highest phenolic content, FRAP and DPPH scavenging activities among the extracts.C. sativum root inhibited DNA damage and prevented MCF-7 cell migration induced by H 2 O 2, suggesting its potential in cancer prevention and inhibition of metastasis. The extract exhibited anticancer activity in MCF-7 cells by affecting antioxidant enzymes possibly leading to H 2 O 2 accumulation, cell cycle arrest at the G 2/M phase and apoptotic cell death by the death receptor and mitochondrial apoptotic pathways. Conclusions: This study is the first report on the antioxidant and anticancer properties of C. sativum root. The herb shows potential in preventing oxidative stress-related diseases and would be useful as supplements used in combination with conventional drugs to enhance the treatment of diseases such as cancer.

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Figure 1 Antioxidant enzyme activities in untreated and treated MCF-7 cells with C. sativum root extract. MCF-7 cells (1.5 × 106) were treated with C. sativum root ethyl acetate extract at 200 μg/ml (IC50 concentration) for 0, 6, 9, 15, 24 and 48 h. DMSO was used to replace the extract in untreated cells. (A) Superoxide dismutase (SOD) activity is expressed in U/ml per 106 cells, while (B) glutathione peroxidase (GPx) and (C) catalase (CAT) activities are expressed in nmol/min/ml per 106 cells. Results are expressed as mean ± SD (n = 3).
Figure 1 Antioxidant enzyme activities in untreated and treated MCF-7 cells with C. sativum root extract. MCF-7 cells (1.5 × 106) were treated with C. sativum root ethyl acetate extract at 200 μg/ml (IC50 concentration) for 0, 6, 9, 15, 24 and 48 h. DMSO was used to replace the extract in untreated cells. (A) Superoxide dismutase (SOD) activity is expressed in U/ml per 106 cells, while (B) glutathione peroxidase (GPx) and (C) catalase (CAT) activities are expressed in nmol/min/ml per 106 cells. Results are expressed as mean ± SD (n = 3).CC BY · PubMed Central
Figure 2 C. Sativum root extract induces cell cycle arrest at G2/M phase in MCF-7 cells. The cell cycle distribution was determined by propidium iodide staining and flow cytometry. MCF-7 cells were treated with the root ethyl acetate extract (276.0 μg/ml) for 24, 48 and 72 h. Untreated cells at each time point were included as controls. The DNA histogram shows the distribution and the percentage of cells in phases of the cell cycle. Results are the mean ± SD of 3 independent experiments. *P < 0.01 compared to the control as tested by the Student’s t-test. Each DNA histogram represents one of the three independent experiments.
Figure 2 C. Sativum root extract induces cell cycle arrest at G2/M phase in MCF-7 cells. The cell cycle distribution was determined by propidium iodide staining and flow cytometry. MCF-7 cells were treated with the root ethyl acetate extract (276.0 μg/ml) for 24, 48 and 72 h. Untreated cells at each time point were included as controls. The DNA histogram shows the distribution and the percentage of cells in phases of the cell cycle. Results are the mean ± SD of 3 independent experiments. *P < 0.01 compared to the control as tested by the Student’s t-test. Each DNA histogram represents one of the three independent experiments.CC BY · PubMed Central
Figure 3 Inhibition of H2O2-induced MCF-7 cell migration by C. sativum root ethyl acetate extract. Results are presented as mean ± SD (n = 3). * P < 0.05 compared to the control (without extract) as tested by the Student’s t-test.
Figure 3 Inhibition of H2O2-induced MCF-7 cell migration by C. sativum root ethyl acetate extract. Results are presented as mean ± SD (n = 3). * P < 0.05 compared to the control (without extract) as tested by the Student’s t-test.CC BY · PubMed Central
Figure 4 HPLC chromatogram of C. sativum root extract. Reverse phase separation was performed at 40°C using a Merck Purospher STAR RP-18 endcapped column (5 μm). The mobile phase consisted of trifluoroacetic acid in water at pH 2.6 (solvent A) and acetonitrile (solvent B). The gradient program consisted of: 0% to 12.5% B for 2.5 min, 12.5% to 100% B for 17.5 min and 100% B for 10 min. The flow rate was kept at 1 ml/min and injection volume was 10 μl. The eluted peaks were monitored at 254 nm. 1: ascorbic acid; 2: p-coumaric acid; 3: butylated hydroxytoluene; S1: unidentified peaks collected for GC-MS analysis. (BSTFA = N,O-bis(trimethylsilyl)trifluoroacetamide).
Figure 4 HPLC chromatogram of C. sativum root extract. Reverse phase separation was performed at 40°C using a Merck Purospher STAR RP-18 endcapped column (5 μm). The mobile phase consisted of trifluoroacetic acid in water at pH 2.6 (solvent A) and acetonitrile (solvent B). The gradient program consisted of: 0% to 12.5% B for 2.5 min, 12.5% to 100% B for 17.5 min and 100% B for 10 min. The flow rate was kept at 1 ml/min and injection volume was 10 μl. The eluted peaks were monitored at 254 nm. 1: ascorbic acid; 2: p-coumaric acid; 3: butylated hydroxytoluene; S1: unidentified peaks collected for GC-MS analysis. (BSTFA = N,O-bis(trimethylsilyl)trifluoroacetamide).CC BY · PubMed Central

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Metadata

Authors
Tang, E. L., Rajarajeswaran, J., Fung, S. Y., & Kanthimathi, M.
Venue
BMC Complementary and Alternative Medicine
Publisher
Springer Science and Business Media LLC
Published
2013-12
Volume
13
Issue
1
ISSN
1472-6882
DOI
10.1186/1472-6882-13-347
PMC
PMC4028854
Cited by
87
References
0
Language
en
Licence
www.springer.com/tdm (unspecified)

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Tang, E. L., Rajarajeswaran, J., Fung, S. Y., & Kanthimathi, M. (2013). Antioxidant activity of Coriandrum sativum and protection against DNA damage and cancer cell migration. BMC Complementary and Alternative Medicine. https://doi.org/10.1186/1472-6882-13-347

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