leukemia is a particular type of cancer characterized by the failure of cell death or disability in differentiation of hematopoietic cells. Chronic myelogenus leukemia (CML) is the most studied kind of this cancer. In this study, an­ti-cancer effect of acetanilide and Para-nitroacetophenone derivatives were investigated in the human leukemia K562 cells. K562 cell line was cultured by initially seeding 1×106 cells per milliliter in RPMI 1640 medium. Cell viability was investigated using trypan blue exclusion and MTT assays.  Cell death in cancer and normal cells was quantified using propidium iodide (PI) and acridine-orange (AO) double staining. The one-way analysis of variance (ANOVA) and the Excel software were used for data analysis. Acetanilide and Para-nitroacetophenone derivatives had strong fatal and concentration-dependent effect on K562 cells and caused cell death mainly through induction of apoptosis. Statistical analysis of cells under fluorescence microscope revealed significant difference in apoptotic cells populations between treated and untreated cells. The results of this investigation clearly indi­cated that Acetanilide and Para-nitroacetophenone derivatives have cytotoxic effects in the K562 cell line. This information revealed also that these compounds may prepare a new thera­peutic approach for the treatment of leukemia.