The biological efeccets of the green coffea on the A549 and the SPC-212 cancer cells

It is preferred that the active ingredients investigated for prevention and treatment of cancer should be organic components rather than being synthetic chemicals. In this study, apoptotic effects of biological activities of green coffee extracts and fractions, previously examined in different aspects, on A549 and SPC-212 cells were investigated. The cytotoxic effects of green coffee extracts and fractions were determined by means of MTT and neutral red dye uptake test and the changes in cell morphology were examined in a fluorescent microscope by using Dual AO/EB staining method and apoptotic index was calculated. The changes occurring in the cell membrane were examined in flow cytometry by doing Annexin V/FITC-PI staining. The activation of the caspase enzyme was measured in Eliza by using caspase-3-8-9 colorimetric kit. IC50 values in A549 cell line were determined as 3 mg/ml in methanol extract; 1, 0.45 and 2.75 mg/ml doses in N-butanol, ethyl acetate and aqueous fractions respectively and 10 mg/ml doses in brewed green coffee. IC50 values in SPC-212 cell line were determined as 0.60 mg/ml in methanol extract; 0.30, 0.28 and 0.80 mg/ml in N-butanol, ethyl acetate and aqueous fractions, respectively and 5 mg/ml IC50 in brewed green coffee. The apoptotic indexes determined according to the morphological images in the ethyl acetate fraction were 25.20 % in SPC-212 and 34.53% in A549. In brewed one, for SPC-212 it was 19.40 % and for A549 it was 36 %. According to the results of Annexin V / FITC-PI staining, in the ethyl acetate fraction and brewing of A549 cells, apoptotic cells were observed in rate of 28.7% and 32.4%, respectively. In SPC-212 cells, apoptotic cells were observed in the rate of 24.7 % in ethyl acetate and 31.2 % in brewing. It is noteworthy that there were approximately 1.5-fold increases in both cells of Caspase-8. It was observed that extract and 3 fractions of green coffee caused reduction and apoptosis in proliferation of A549 and SPC-212 cells. It was determined that in both cell lines, the effect of acetate fraction and brewed green coffee to apoptosis was higher than the others. When comparing the two cell lines, it was found that SPC-212 cells were more sensitive than A549 cells in terms of cytotoxic and apoptotic effects. Keywords: Cytotoxicity, A549, SPC 212, Green Tea.