Generating Chemoresistant Breast Cancer Cell Lines

Chemoresistant BT-474 and BT-474 NRP-1 variant cell lines were generated in a similar protocol to that described previously [13 (link)]. Briefly, the cells were treated with four cycles of a combination of 0.5 uM of Doxorubicin (Brand name Adriamycin, Pharmacia, Italy) + 300 nM Cyclophosphamide (Brand name Cytoxan, Baxter, Germany) (cells will be referred to as 4xAC) followed by four cycles of 20 nM Paclitaxel (Brand name Taxol, EBEWE Pharma, Austria) (cells will be referred to as 4xAC + 4xPAC). Each cycle was for a duration of 72 h followed by a recovery period until confluency was achieved prior to commencement of the next cycle. Protein lysate and RNA was extracted from the 4xAC and 4xAC + 4xPAC resistant cell lines and stored at − 80 °C.