Expression of Her2 and drug efflux proteins in parental and resistant cell models. Expression of Her2 and drug efflux proteins in parental and resistant.

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Expression of Her2 and drug efflux proteins in parental and resistant cell models. Expression of Her2 and drug efflux proteins in parental and resistant cell models. A, immunoblot analyses of 80 μg total protein from whole-cell lysates show Her2 levels decreased in JIMT1-TM cells and partially reduced in 361-TM cells. ABCC1 (MRP1) expression is elevated in 361-TM cells, but not in JIMT1-TM cells or in their parental counterparts. A549 and H69AR cells serve as positive controls for ABCC1. No detectable ABCB1 (MDR1) was observed in either 361-TM or JIMT1-TM cells, compared with positive controls KB-8.5 and KB-V1 cells. Cell lines were analyzed by flow cytometry for trastuzumab antibody binding (B) and internalization (C). 361, blue lines; JIMT1, red lines; parental cells, open symbols; TM-ADC–resistant cells, closed symbols. D, flow cytometric analysis of ABCC1 antibody shows elevated binding in 361-TM cells relative to parental 361 cells. Secondary antibody only (gray bar); 10 μg/mL ABCC1 antibody (blue bar). E, efflux study demonstrates increased accumulation of rhodamine in 361-TM cells compared with parental cells in the presence of reversan. *, P < 0.001 difference compared with 361 cells as evaluated by Student t test. Reference cell lines A549 and H69AR express ABCC1. Frank Loganzo et al. Mol Cancer Ther 2015;14:952-963 ©2015 by American Association for Cancer Research