Evaluation of anti-proliferative activity of Eryngium caucasicum on cancerous (B16) and non- cancerous (HFFF-2) cell lines in vitro

Abstract

Introduction: Eryngium , a member of the Apiaceae family, have shown different pharmacological effects including anti-cancer, anti-oxidant effects. Previous reports on the anti-cancer activities of Eryngium genus and Eryngium caucasicum in some cancer cell lines has led us to study the anti-proliferative activity of Eryngium caucasicum on B16 cell line. Objective: Evaluation of anti-proliferative activity of Eryngium caucasicum in B16 cells lines and non- cancerous cells, HFFF-2, in vitro. Methods: Air dried powder of E. caucasicum was soxhelet extracted using n-Hexane, dichloromethane and methanol as solvents. MTT assay was used to assess the cytotoxic effects of the extracts on B16 and HFFF-2 cell lines. The fractions of the most potent anti-cancer effects extract, was prepared using vacuum liquid chromatography, then the growth inhibitory effects of the fractions were evaluated using MTT assay. Annexin V/PI assay was used to study the induction apoptosis by the fractions in cancer cells. Results: N-Hexane extract showed the most potent cytotoxic effects in B16 cell line and it was used for fractioning. 40% and 60% VlC fractions of n-Hexane extract shows the most potent cytotoxic effect against B16 cells (p<0.001). Annexin V/PI analysis revealed that while cell death caused by n-Hexane is through the induction of both apoptosis and necrosis, 40% and 60% VlC fractions of n-Hexane extract mostly induce apoptosis in B16 cells. Importantly 40% and 60% VlC fractions didn’t show significant cytotoxic effects in normal cells when it was used at IC50 concentration. Conclusion: n-Hexane extract of Eryngium caucasicum and its 40% and 60% fractions show the most cytotoxic effects against B16 cell line as compared with other extracts and the control groups. Cell death caused by 40% and 60% fractions is mostly through induction of apoptosis whereas the extracts cause cells death through both apoptosis and necrosis.