Cytotoxicity of Fenugreek Sprout and Seed Extracts and Their Bioactive Constituents on MCF-7 Breast Cancer Cells

Abstract

Trigonella foenum-graecum L. (fenugreek), a member of the legume family (Fabaceae), is a promising source of bioactive phytochemicals, which explains its traditional use for a variety of metabolic disorders including cancer. The current study aimed to evaluate extracts of fenugreek seeds and sprouts, and some of their constituents, to compare their cytotoxic and antiproliferative activities in MCF-7 breast cancer cells. The extracts were chemically characterised using high-resolution accurate mass liquid chromatography-mass spectrometry to reveal the detection of compounds assigned as flavone C-glycosides including those derived from apigenin and luteolin, in addition to isoflavones. Five different flavones or their glycosides (apigenin, vicenin-2, vitexin, luteolin and orientin) and two isoflavones (daidzein and formononetin) were quantified in the fenugreek extracts. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay using MCF-7 cells treated with fenugreek methanolic extracts showed dose- and time-dependent effects on cell viability. The MCF-7 cancer cells treated with the fenugreek methanolic extracts also displayed increased relative mitochondrial DNA damage as well as suppressed metastasis and proliferation. This study demonstrates the potential anti-cancer effects of fenugreek seeds and sprouts and reveals fenugreek sprouts as an untapped resource for bioactive compounds.

Keywords: MCF-7; cancer; cytotoxicity; fenugreek; iron.

Figure 1

Cell viability of MCF-7 cells treated with various concentrations of fenugreek sprout and seed methanol extracts (0–6000 µg/mL) for 24 h. 1st Fenugreek Sprouts Methanol Extraction [1st FPME], 3rd Fenugreek Sprouts Methanol Extraction [3rd FPME], 1st Fenugreek Seeds Methanol Extraction [1st FSME], 3rd Fenugreek Seeds Methanol Extraction [3rd FSME]. The values are presented as means n = 6 ± SEM.

Figure 2

Fenugreek-inhibited cell growth in MCF-7 human breast cancer cells. The viability of the cells was analysed using cell MTT assays. 1st Fenugreek Sprouts Methanol Extraction [1st FPME], 3rd Fenugreek Sprouts Methanol Extraction [3rd FPME], 1st Fenugreek Seeds Methanol Extraction [1st FSME], 3rd Fenugreek Seeds Methanol Extraction [3rd FSME]. The IC₅₀ treatments of the first FSME at 5605 µg/mL, first FPME at 812 µg/mL, third FSME at 1400 µg/mL, and third FPME at 526 µg/mL for 24, 48, and 72 h decreased the viability of the MCF-7 cells in a time-dependent manner. The values are presented as means n = 6 ± SEM. The data were analysed using a one-way ANOVA test. Different asterisks indicate significant differences between data in the same group (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001).

Figure 3

Fenugreek-inhibited cell proliferation of MCF-7 human breast cancer cells. The proliferation of cells was analysed using a cell proliferation assay. 1st Fenugreek Sprouts Methanol Extraction [1st FPME], 3rd Fenugreek Sprouts Methanol Extraction [3rd FPME], 1st Fenugreek Seeds Methanol Extraction [1st FSME], 3rd Fenugreek Seeds Methanol Extraction [3rd FSME]. The IC₅₀ treatment of the 3rd FSME at 5605 µg/mL, 1st FPME at 812 µg/mL, 3rd FSME at 1400 µg/mL, and 3rd FPME at 526 µg/mL for 24, 48, and 72 h decreased the proliferation of the MCF-7 cells in a time-dependent manner. The values are presented as means n = 6 ± SEM. The data were analysed using a one-way ANOVA test. Different asterisks indicate significant differences between data in the same group (** p ≤ 0.01, and **** p ≤ 0.0001).

Figure 4

The mtDNA relative content in the MCF-7 cells. Amplified by RT-PCR and normalised to the human Beta-2 microglobulin [hB2M] nuclear gene after IC₅₀ treatment of the 3rd FSME at 5605 µg/mL, 1st FPME at 812 µg/mL, 3rd FSME at 1400 µg/mL, and 3rd FPME at 526 µg/mL in the MCF-7 cells for 24 h; H₂O₂ was used as a positive control. 1st Fenugreek Sprouts Methanol Extraction [1st FPME], 3rd Fenugreek Sprouts Methanol Extraction [3rd FPME], 1st Fenugreek Seeds Methanol Extraction [1st FSME], 3rd Fenugreek Seeds Methanol Extraction [3rd FSME], hydrogen peroxide [H₂O₂] (Sigma H1009). The values are presented as means n = 4 ± SEM. The data were analysed using a two-way ANOVA test. The different letters on top of the bars indicate significant differences (p ≤ 0.05) between the data.