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  • Fenugreek extract as an inducer of cellular death via autophagy in human T lymphoma Jurkat cells.

Fenugreek extract as an inducer of cellular death via autophagy in human T lymphoma Jurkat cells.

BMC complementary and alternative medicine (2012-11-01)
Nasser M Al-Daghri, Majed S Alokail, Khalid M Alkharfy, Abdul Khader Mohammed, Sherif H Abd-Alrahman, Sobhy M Yakout, Osama E Amer, Soundararajan Krishnaswamy
ABSTRACT

Drugs used both in classical chemotherapy and the more recent targeted therapy do not have cancer cell specificity and, hence, cause severe systemic side effects. Tumors also develop resistance to such drugs due to heterogeneity of cell types and clonal selection. Several traditional dietary ingredients from plants, on the other hand, have been shown to act on multiple targets/pathways, and may overcome drug resistance. The dietary agents are safe and readily available. However, application of plant components for cancer treatment/prevention requires better understanding of anticancer functions and elucidation of their mechanisms of action. The current study focuses on the anticancer properties of fenugreek, a herb with proven anti-diabetic, antitumor and immune-stimulating functions. Jurkat cells were incubated with 30 to 1500 μg/mL concentrations of 50% ethanolic extract of dry fenugreek seeds and were followed for changes in viability (trypan blue assay), morphology (microscopic examination) and autophagic marker LC3 transcript level (RT-PCR). Incubation of Jurkat cells with fenugreek extract at concentrations ranging from 30 to 1500 μg/mL for up to 3 days resulted in cell death in a dose- and time-dependent manner. Jurkat cell death was preceded by the appearance of multiple large vacuoles, which coincided with transcriptional up-regulation of LC3. GC-MS analysis of fenugreek extract indicated the presence of several compounds with anticancer properties, including gingerol (4.82%), cedrene (2.91%), zingerone (16.5%), vanillin (1.52%) and eugenol (1.25%). Distinct morphological changes involving appearance of large vacuoles, membrane disintegration and increased expression of LC3 transcripts indicated that fenugreek extract induced autophagy and autophagy-associated death of Jurkat cells. In addition to the already known apoptotic activation, induction of autophagy may be an additional mechanism underlying the anticancer properties of fenugreek. This is the first report showing fenugreek as an inducer of autophagy in human cells and further work is needed to define the various intermediates of the autophagic pathway.

MATERIALS
Product Number
Brand
Product Description

Supelco
Mettler-Toledo Calibration substance ME 51143093, Vanillin, traceable to primary standards (LGC)
Sigma-Aldrich
Vanillin, ≥97%, FCC, FG
Sigma-Aldrich
Vanillin, natural, ≥97%, FCC, FG
Sigma-Aldrich
Vanillin, ReagentPlus®, 99%
Sigma-Aldrich
Vanillin, tested according to Ph. Eur.
Sigma-Aldrich
Guaiacol, oxidation indicator
Sigma-Aldrich
Guaiacol, natural, ≥99%, FG
Supelco
Guaiacol, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Vanillin melting point standard, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Vanillin, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Vanillylacetone, ≥96%, FG