Bioflavonoids are a broadly distributed class of plant pigments, universally present in vascular plants and responsible for much of the coloring in nature. They are strong antioxidants that occur naturally in foods and can inhibit carcinogenesis. The consumption of food containing high amount of bioflavonoids has been reported to lower the risk of various cancers. The aim of our study was to investigate the anti-cancer effects of bioflavonoids Acacetin (ACA) and Luteolin (LUT) in cultured HO-1 Human Melanoma Cancer Cells. We conducted cell proliferation and cell viability studies using trypan blue cell exclusion method, Western blot analysis, DNA fragmentation using agarose Gel electrophoresis in HO-1 melanoma cells treated with bioflavonoids ACA and LUT, alone and in combination. We also observed the changes of cell morphologies after treating the cells with these biofalvonoids. Our data shows that bioflavonoids significantly inhibited the growth of melanoma cells and induced apoptosis, which was determined by observing a time-dependent up-regulation in expression of the proapoptotic Bad, p-53 & Apaf-1 proteins and time-dependent down-regulation of anti-apoptotic Bcl-xL protein expressions in cells treated from 24 to 72 hours. Maximum expressions were observed after 72 hours treatment for the pro-apoptotic genes. We confirmed the occurrence of apoptosis by performing DNA fragmentation experiments. In summary, these studies demonstrate that ACA and LUT alone and in combination significantly inhibit cell proliferation of HO-1 cells by inducing apoptosis. Synergistic effects were observed while the cells were treated with the combination of ACA and LUT. The induction of apoptosis by Acacetin and Luteolin may offer a pivotal mechanism for its cancer therapeutic and chemo-preventive action. Further studies are required to investigate the detailed mechanism of apoptosis in response to treatment of HO-1 cells with these flavonoids and other bioflavonoids combination.