Combination therapy with a new and active component has been illustrated as a strategy in breast cancer treatment versus the conventional methods. Therefore, the current research investigated the role of quinacrine (QC) in enhancing the efficacy of doxorubicin (DOX) in MDA-MB-231 cancer cells. Anti-proliferation effect of co-administration of QC and DOX was examined using MTT assay and DAPI staining analysis. FACS techniques were performed to identify the molecular mechanisms of apoptosis and cell cycle progression. Moreover, the transwell migration assay was applied to measure the role of QC in the invasiveness of MDA-MB-231cells; the results showed that by applying QC along with DOX, the migration rate of cancer cells decreased significantly (2.41 fold; p < 0.5). The 1050 values were 1.2 +/- 0.1 and 4.4 +/- 0.4 mu M for DOX and QC, respectively. The population of apoptotic cells was increased from 35 +/- 2% to 40 +/- 4% (p < 0.05) under treatment with DOX and QC. QC triggered mRNA level of pro-apoptotic marker Bax, and decreased anti-oxidant marker Nrf2 and anti-apoptotic marker bcl-xl, cyclin-B1 expression in MDA-MB-231cells. We found the combination therapy with DOX and QC a novel therapeutic approach that could enhance the efficacy of chemotherapy.