Background: Ovarian cancer remains a major therapeutic challenge because of late diagnosis, recurrence, and treatment resistance. Fenugreek (Trigonella foenum-graecum L.) contains several bioactive constituents with potential anticancer properties. This study investigated the cytotoxic effect of aqueous fenugreek seed extract on SKOV-3 human ovarian cancer cells and evaluated its association with the expression of apoptosis-related genes.
Methods: SKOV-3 cells were treated with aqueous fenugreek seed extract at concentrations of 1, 10, 50, 100, and 200 µg/mL for 24 and 48 h. Cell viability was assessed using the MTT assay, and half-maximal inhibitory concentrations (IC50) were determined by nonlinear regression. To investigate apoptosis-related molecular changes, cells were treated with the experimentally determined IC50 concentration, and the relative mRNA expression levels of BAX, BCL2, and CASP3 were evaluated by quantitative real-time PCR (qRT-PCR) using GAPDH as the reference gene. Relative gene expression was calculated using the 2^−ΔΔCt method.
Results: Aqueous fenugreek extract reduced SKOV-3 cell viability in a concentration- and time-dependent manner. The estimated IC50 values were 38.5 µg/mL at 24 h and 19.7 µg/mL at 48 h, indicating greater cytotoxicity with prolonged exposure. Treatment at the IC50 concentration significantly increased BAX expression to 2.83-fold and CASP3 expression to 3.21-fold, while BCL2 expression decreased to 0.43-fold relative to untreated cells. Consequently, the BAX/BCL2 expression ratio increased approximately 6.6-fold, indicating a shift toward a pro-apoptotic molecular profile.
Conclusion: Aqueous T. foenum-graecum seed extract exerted concentration- and time-dependent cytotoxic effects on SKOV-3 ovarian cancer cells and was associated with coordinated modulation of apoptosis-related genes, characterized by increased BAX and CASP3 expression, decreased BCL2 expression, and an elevated BAX/BCL2 ratio. These findings suggest that modulation of apoptosis-related signaling may contribute to the cytotoxic activity of aqueous fenugreek extract. Further functional and protein-level studies are required to confirm apoptosis and clarify the underlying molecular mechanisms.