In the systematic screening programme for cytotoxic compound from marine actinomycetes, the compound furan-2-yl acetate (F2A) from Streptomyces VITSDK1 spp. The structure of the compound was unequivocally determined by spectral studies. It was previously found that F2A has potential antiviral activity against fish nodavirus. In the present study, the cytotoxicity of F2A was studied using (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay which showed the IC50 values were less than 15 μg mL-1 against various tumor cell lines, whereas it was >25 μg mL-1 against non-tumor cell lines. F2A inhibited the cell proliferation in a dose-and time-dependent manner. Furthermore, the cytotoxic mechanism was determined in HeLa cells. The morphological analysis, Hoechst staining and DNA fragmentation studies revealed the apoptosis mediated cell death. The cytosolic protein analysis of F2A treated HeLa cells by immunoblotting showed the mitochondrial cytochrome c release, increased expression of caspase 3 and caspase 9 with PARP cleavage. There was no change in the caspase-8 levels. The Bcl-2 was found to be down regulated and Bax was up regulated in the F2A treated cells. Further, the apoptosis induction and cell death was found to be mediated by Reactive Oxygen Species (ROS) and lipid peroxidation. A molecular docking study of F2A with 28 selected cancer drug target enzymes provides some insight on mode of activity of the compound. The findings showed that the F2A exhibits selective cytotoxicity towards tumor cells at a lower concentration via apoptosis. © 2013 Academic Journals Inc.