InfIuence of BCRP overexpression on proIiferation and apoptosis of human breast cancer ceIIs

YU Zhaojin; YU Jiankun; YU Lifeng; WEI Minjie; ZHAO Lin

doi:10.3969/j.issn.1002-266X.2016.25.006

Abstract：Objective To investigate the effect of breast cancer resistance protein (BCRP)overexpression on cell proliferation and apoptosis of breast cancer cells MDA-MB-435S.Methods MDA-MB-435S cells in the logarithmic phase were randomly divided into three groups:the positive transfection group,in which the eukaryotic expression plasmid pcD-NA3.1 /BCRP was identified by restriction analysis and sequencing and then was transfected into MDA-MB-435S cells,the positive control group,in which the cells were transfected with 2 μg pcDNA3.1 empty plasmid at the same time,and the blank control group,in which the cells were cultured with 2 mL nutrient solution.The mRNA and protein expression of BCRP was determined by RT-PCR and Western blotting,respectively.After 48-hour transfection,7 mitomycin (MMC) concentration groups (0.01,0.10,2.50,5.00,10.00,20.00,40.00 μmol/L)were set up in each group,each con-centration group had 3 complex holes which were added with 10 μL MMC.Cell counting kit-8 (CCK-8)and flow cytometry were used to determine the influence of BCRP on cell proliferation and apoptosis of MDA-MB-435S cells,respectively.Re-sults Restriction analysis and sequencing proved that recombinant plasmid pcDNA3.1 /BCRP was constructed correctly. Both the expression of BCRP mRNA and protein was up-regulated in MDA-MB-435S cells after transfection 24,48 or 72 h in the positive transfection group (P <0.05).No significant difference was found between the positive transfection group and the negative control group (P >0.05).After MMC treatment of 24 hours,CCK-8 showed that the cell survival rate in the transfection group was significantly higher than that of the negative control group (P <0.05).Conclusion BCRP overexpression promotes the proliferation of MDA-MB-435S cells,but has no significant effect on apoptosis.