Apoptotic DNA fragmentation trİggered by combination theraphy of 5-FU and CAPE in 4549 cell line

Abstract

Non-small cell lung cancer cell (NSCL)is a leading cause oF caNcer mortality over the World. Caffeic Acid Phenethyl Ester (CAPE) is a major active component in propolis. It has been previorısly identified as a strong antıoxicJant, antiinflammatory, antiviral and anticancer molecule. we aiıned to investigate the comparative effects of 5-FU, CAFE with single and combine treatment in A549 cells. We investigaıe to analysis of apoptosis tıy DNA fragmentation in A549 cells. Thus, we further exarıined DNA fragmentation to clarify whether CAPE analogues induced apoptosis or not. Cells were cullured in RPlVII-1640 in a humidified atmosphere of 5%CO2 at 37C. Cell viability was determined by MTT assay, The lC50 values were detected for 5-FU, CAPE, and combined treatment by 50uM, 4uM and 12,5uM +1 uM respectively. We compared the effect of monotherapy and polytherapy of drugs on cells. cells were treated with determined concentration for 24 and 48 hours. After treatment, cells were isolated according to DNA fragmentation protocol and DNA fragments showed on 3% agarose gel. For cell viability, cells were treated with lC50 value for each drug and combination 24h, 48h of incubation. combine therapy is more effective than single therapy of these drugs. We determined that DNA fragmentation, a marker for induction of apoptosis, increased witlh 5-FU treatment at 48 hours, Tliese results suggest that 5-FU is more effectiective than CAPE to induction of apoptosis. This study is a basic qualitative study for the investigate of the apoptosis pathway triggered by 5-FU.