title: Proteome analysis of multidrug-resistant, breast cancer-derived microparticles.
abstract: Cancer multidrug resistance (MDR) occurs when cancer cells evade the cytotoxic actions of chemotherapeutics through the active efflux of drugs from within the cells. Our group have previously demonstrated that multidrug-resistant breast cancer cells spontaneously shed microparticles (MPs) and that these MPs can transfer resistance to drug-responsive cells and confer MDR on those cells in as little as 4 h. Furthermore, we also showed that, unlike MPs derived from leukaemia cells, breast cancer-derived MPs display a tissue selectivity in the transfer of P-glycoprotein (P-gp), transferring the resistance protein only to malignant breast cells. This study aims to define the proteome of breast cancer-derived MPs in order to understand the differences in protein profiles between those shed from drug-resistant versus drug-sensitive breast cancer cells. In doing so, we detail the protein cargo required for the intercellular transfer of MDR to drug-sensitive recipient cells and the factors governing the transfer selectivity to malignant breast cells. We describe the first proteomic analysis of MPs derived from human breast cancer cells using SDS PAGE and liquid chromatography-tandem mass spectrometry (LC/MS/MS), in which we identify 120 unique proteins found only in drug-resistant, breast cancer-derived MPs. Our results demonstrate that the MP-mediated transfer of P-gp to recipient cells occurs alongside CD44; the Ezrin, Radixin and Moesin protein family (ERM); and cytoskeleton motor proteins within the MP cargo.
authors.affiliation: Discipline of Pharmacy, The Graduate School of Health, University of Technology, Sydney, Australia.
authors.forename: Deep
authors.initials: D
authors.lastname: Pokharel
authors.affiliation: The ithree Institute, University of Technology, Sydney, Australia ; Proteomics Core Facility, University of Technology, Sydney, Australia.
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Padula
authors.affiliation: Discipline of Pharmacy, The Graduate School of Health, University of Technology, Sydney, Australia.
authors.forename: Jamie F
authors.initials: JF
authors.lastname: Lu
authors.affiliation: The ithree Institute, University of Technology, Sydney, Australia.
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Tacchi
authors.affiliation: Discipline of Pharmacy, The Graduate School of Health, University of Technology, Sydney, Australia.
authors.forename: Frederick
authors.initials: F
authors.lastname: Luk
authors.affiliation: The ithree Institute, University of Technology, Sydney, Australia ; Proteomics Core Facility, University of Technology, Sydney, Australia.
authors.forename: Steven P
authors.initials: SP
authors.lastname: Djordjevic
authors.affiliation: Discipline of Pharmacy, The Graduate School of Health, University of Technology, Sydney, Australia.
authors.forename: Mary
authors.initials: M
authors.lastname: Bebawy
keywords: P-glycoprotein; breast cancer; liquid chromatography–tandem mass spectrometry; microparticles; multidrug resistance; proteomics; tissue selectivity