Effects of Fenugreek on Lung Cancer / In Vitro Study

Abstract

Lung cancer is the leading cause (account for 18%) of cancer death in both men and women world-wide.The overall 5-year survival rate for all stages combined is disappointing (15%).The aim of this study is to evaluate the possible cytotoxic effects of fenugreek on lung cancer cell line and to determine its IC50 alone and in combination with cisplatin. And to study the effects of fenugreek on the expression of each of p53 and EGFR QU-DB lung cancer cells were cultured in Eagle's MEM culture media supplemented with 5% FBS and antibiotics. The cells were seeded in 96 well plate and the cytotoxic effects of each of cisplatin [25-0.195 µl/ml (or µg/ml)] and fenugreek [300-1.1719 µl/ml (each one µl is extracted from 25 µg of dried seed)] was determined using neutral red uptake (NRU) assay for 24, 48, and 72 hours in comparison with their corresponding control groups. Combined effect of each of fenugreek and cisplatin was determined also using NRU assay. Cytotoxicity was further assessed by trypan blue exclusion assay at IC50 of each agent for 48 hours duration. Immunocytochemistry assay was performed also to detect EGFR and p53 expression. Cisplatin induced a directly proportional, dose-dependent and time-dependant cytotoxic effect with an IC50 of 8.5 µg/ml and 7.3 µg/ml after 48 hrs and 72 hrs of exposure respectively. Significant differences (p<0.05) were observed in optic density of cisplatin group from that of the control for all tested concentrations.Fenugreek extract also induced a directly proportional, dose-dependent and time-dependant cytotoxic effect in experiments with 48 hrs and 72 hrs of exposure with an IC50 of 88.25 µl/ml and 125 µl/ml respectively (each one µl is extracted from 25 µg of dried seed). While it produces a growth enhancing effect in 24 hrs exposure experiment. Significant differences (p<0.05) were observed in optic density of fenugreek from that of the control at concentrations of 37.5 µl/ml and above.Fenugreek produces an antagonistic action when combined with cisplatin, combination index (CI) >1.3.Cisplatin highly significantly (p<0.005) increased EGFR expression at different concentrations. While fenugreek extract highly significantly (p<0.005) reduced EGFR expression at 300 µl/ml (each one µl is extracted from 25 µg of dried seed).Cisplatinand fenugreek highly significantly (p<0.005) decreased the expression of P53.Monotherapy of fenugreek have anticancer effect on lung cancer cell line, but an antagonizing effect to cisplatin when combined with it.Fenugreek may have a beneficial therapeutic effect in decreasing EGFR expression and decreasing mutant p53 expression. Further study is recommended to explore the effect of fenugreek on other cell cycle proteins and to study their potential beneficial therapeutic effects in vivo.