Chemotherapy is one of various cancer therapeutic treatments that usually has efficacy in cancer treatment, but it has many toxic or side effects on other normal tissues or cells. The present study involves the reduction of some toxic effects, such as cytotoxicity and DNA damage in normal human skin melanocytes (HFB4) that was exposed to doxorubicin (Dox) as a chemotherapeutic drug. HFB4 cells were treated by Dox alone and Dox which was combined with vitamin C (Vit. C). The MTT technique, alkaline comet assay as well as biochemical parameters were used. The results showed that Dox treatment alone at 0.26 and 0.39 µg/ml caused oxidative stress and DNA damage as well as caused the reduction of superoxide dismutase activity and lipid peroxidation. However, addition of Vit. C at concentrations of 32, 65, and 130 µg/ml reduced these effects on HFB4 cells. Also, it increased the proliferation of HFB4 cells and decreased the DNA breaks. Further, the addition of Vit. C with Dox reduced the lipid peroxidation and enhanced the anticancer activity of Dox. Conclusion: The combination of natural compounds (Vit. C) at low concentrations with Dox may be able to protect normal cells from genotoxicity and may be enhanced the anticancer activity of Dox against cancer cells.