In vitro evaluation of cytotoxic activity of Trigonella foenum-graecum on human cervical cancer HeLa cells

Abstract

Trigonella foenum-graecum L. (fenugreek) is a plant of the Fabaceae family. The species contains steroidal saponins, flavonoids, polysaccharides and phenolic acids, which are pharmacological active compounds with anti-inflammatory, antioxidant and hypoglycemic properties. This study reports on the evaluation of cytotoxic effects of the plant methanolic and aqueous extracts, flavonoid fraction and isolated flavonoids -orientin, isoorientin, vicenin-1, -2, and -3- on human cervical adenocarcinoma cell line HeLa. The antiproliferative activity was estimated on the cells with MTT assay and the xCELLigence system (Roche), which allows continuous monitoring of cell viability. The obtained data show that both methanolic and aqueous extracts, as well as the flavonoid fraction have significant cytotoxic activity on the cells with IC50 values of 13.47 ± 0.62 µg/mL, 17.43 ± 0.30 µg/mL and 3.91 ± 0.03 µg/mL, respectively. The flavonoids did not exhibit activity on the cells (IC50> 50 µg/mL). Additionally, HeLa cells treated with the fraction were stained with annexin V and analyzed on the Muse Cell Analyzer (Merck Millipore). The flow cytometry analysis shows that the fraction induces apoptosis in the tested cells in a dose-dependent manner. To confirm induction of apoptosis in HeLa cells incubated with the fraction, we stained the cells with Hoechst 33342 dye and observed the chromatin condensation and nuclear fragmentation in the cells.