A steroidal derivative from Trigonella foenum graecum L. that induces apoptosis in vitro and in vivo

Trigonella foenum graecum L. is a dietary herb used in traditional medicine system. In this study, we investigated the cytotoxicity, antitumor, antimetastatic and antiangiogenic effect of the steroidal compound, ethyl iso-allocholate isolated from T. foenum graecum L. seeds against A549 lung cancer cells in vitro and in vivo. Among all the isolated compounds, the ethyl iso-allocholate rendered the highest cytotoxicity potential. It showed least percentage cell viability in trypan blue assay and lowest nuclei count in hoechst staining. The caspase glo assay and western blot analysis showed the significant caspase 3 cleavage, indicating caspase dependent apoptosis. Consistent with the in vitro data, ethyl iso-allocholate showed highest percentage tumor growth inhibition i.e. 80 ± 5% in zebrafish, equivalent to doxorubicin. It significantly reduced angiogenesis to 5 ± 0.8% (**P < 0.01), compared to negative control group which was 60 ± 2%. The ethyl iso-allocholate showed 55 ± 3% inhibition in liver metastasis. To investigate the safety of the compounds on normal tissues, the percentage mortality was examined. The ethyl iso-allocholate showed zero percent mortality of zebrafish. These results indicate that the steroidal derivative isolated from T. foenum-graecum seeds induces caspase dependent apoptosis in cancer cells and reduces tumor growth, metastasis and angiogenesis in vivo, as well as it is safe on the normal tissues. The in vitro and in vivo anticancer studies suggest that the cytotoxic compound ethyl iso-allocholate has potential application in pharmaceutical industry.