Eryngium billardieri Extract and Fractions Induce Apoptosis in Cancerous Cells

Abstract

Background: Eryngium is a genus flowering plant in the Umbelliferae family, having pharmacological properties, such as anti-inflammatory and anti-diabetic. Given the nature of melanoma and breast cancers in recent years and the fact that the anti-cancer properties of Eryngium billardieri on mentioned cell lines have not been studied, the present study was conducted to explore these properties.

Objective: The mechanisms of cytotoxicity and apoptosis of aerial parts of various extracts and fractions of E. billardieri on cancerous cells and normal cells were investigated.

Methods: Samples were collected from natural habitats, dried and then extracted by Soxhlet apparatus with solvents of n-Hex, DCM and methanol, respectively. The cytotoxic effects of the extracts were investigated by the MTT method on MCF7, B16 and HFF-2 classes for 24 and 48 hours. Flow cytometry was also used to investigate the mechanism of cytotoxicity, and it has been confirmed by Real-time PCR of p53 and Bax genes as to which comprise the apoptosis regulatory proteins. Meanwhile, volatile compounds of extracts were identified by the GC-MS method.

Results: The obtained data showed that the n-Hex extract of E. billardieri on B-16 and MCF7 cell lines and dichromethane extract on MCF7 cell line had the most significant cytotoxic effect compared to DMSO control (p-value <0.001). Our finding showed that the mechanism of n-Hex extract with 80% and 100% vlc fractions on B16 induced apoptotis compared to HFF-2 control cells; moreover, n-Hex extract and 80% vlc fraction on MCF7 were apoptotic. The major compounds of n-Hex, DCM, and 80% and 100% fractions of n-Hex extract obtained from GC-MS were non-terpenoid.

Conclusion: Non-terpenoid compounds of E. billardieri can be responsible for exhibiting the most cytotoxic effects on MCF7 and B16 cell lines with apoptotic mechanism, and n-Hex extract was found to have the most significant inhibitory effect on cancerous cells compared to the HFF-2 control cells by employing the mechanism of apoptosis.

Keywords: Eryngium billardieri; GC-MS; apoptosis; cancer; cytotoxic effects; flow cytometry.