Abstract：OBJECTIVE To establish a doxorubicin hydrochloride(DOX) resistant cell line model of human triple-negative breast cancer MDA-MB-231/DOX and investigate its biological characteristics. METHODS Doxorubicin hydrochloride-resistant breast cancer cell line was established by continuous induction with a gradual increasing dose of DOX. The cell morphology changes were observed under an optical microscope; the half maximal inhibitory concentration(IC50) and RI(resistance index) of drug resistant cells to chemotherapeutics were examined by cytotoxicity experiments and the cell growth curve was plotted; the cell cycle distribution was determined by flow cytometry; the protein expression of P-glycoprotein(P-gp), multidrug resistance-associated protein(MRP2), breast cancer resistance protein(BCRP) and glucose transporter(GLUT1, GLUT3) in cells were examined by Western blot; the biological characteristics of MDA-MB-231/DOX cells in the aspect of energy metabolism were investigated by detecting the concentration of pyruvic acid, lactic acid and adenosine triphosphate(ATP) in cells. RESULTS The established MDA-MB-231/DOX resistant cell line was resistant to different concentrations of DOX, and its resistance index to DOX was in the range of 61-189. The resistance indexes of MDA-MB-231/DOX cells to paclitaxel(PTX), cisplatin(CDDP) and 10-hydroxycamptothecin(HCPT) were 1 585.12, 29.67 and 2.01, respectively; the protein expressions of P-gp and MRP2 in drug-resistant MDA-MB-231/DOX cells were significantly higher than those in parental MDA-MB-231 cells; the concentrations of pyruvic acid, lactic acid and ATP, and the protein expression of GLUT1 in MDA-MB-231/DOX cells were also higher than that in MDA-MB-231 cells. CONCLUSION The human triple-negative breast cancer resistant cell line MDA-MB-231/DOX was successfully established. The cell line exhibits multiple drug resistanc and its biological characteristics are different from the parental cells to some extent. The present research will provide some experimental basis for establishing breast cancer model in vitro and exploring the related drug resistance mechanism.
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