Detection and characterisation of multi-drug resistance protein 1 (MRP-1) in human mitochondria.

British Journal of Cancer
E A Roundhill, S A Burchill

Abstract

Overexpression of plasma membrane multi-drug resistance protein 1 (MRP-1) can lead to multidrug resistance. In this study, we describe for the first time the expression of mitochondrial MRP-1 in untreated human normal and cancer cells and tissues. MRP-1 expression and subcellular localisation in normal and cancer cells and tissues was examined by differential centrifugation and western blotting, and immunofluorescence microscopy. Viable mitochondria were isolated and MRP-1 efflux activity measured using the calcein-AM functional assay. MRP-1 expression was increased using retroviral infection and specific overexpression confirmed by RNA array. Cell viability was determined by trypan blue exclusion and annexin V-propidium iodide labelling of cells. MRP-1 was detected in the mitochondria of cancer and normal cells and tissues. The efflux activity of mitochondrial MRP-1 was more efficient (55-64%) than that of plasma membrane MRP-1 (11-22%; P<0.001). Induced MRP-1 expression resulted in a preferential increase in mitochondrial MRP-1, suggesting selective targeting to this organelle. Treatment with a non-lethal concentration of doxorubicin (0.85 nM, 8 h) increased mitochondrial and plasma membrane MRP-1, increasing resistance to MR...Continue Reading

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Citations

Nov 7, 2015·Drug Metabolism Reviews·Jamie F LuMary Bebawy
May 22, 2013·Proceedings of the National Academy of Sciences of the United States of America·Julien MarcouxCarol V Robinson
Feb 20, 2017·Biochimica Et Biophysica Acta. General Subjects·Julie DartierJean-François Dumas
Mar 16, 2013·The Anatomical Record : Advances in Integrative Anatomy and Evolutionary Biology·Kai TangLi-Ming Li
Jan 2, 2016·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Elizabeth RoundhillSusan Burchill
Aug 16, 2014·Asian Pacific Journal of Cancer Prevention : APJCP·Jin-Peng HeJie-Xiong Feng
Sep 26, 2020·Cell Death & Disease·Rui LiuYuanyan Liu
Oct 8, 2014·Experimental and Therapeutic Medicine·Xiao-Hong XieXi-Dong Gu
Dec 17, 2019·Trends in Biochemical Sciences·Robert C FordJohn Schuetz
May 12, 2018·Phytomedicine : International Journal of Phytotherapy and Phytopharmacology·Zhaolin ChenAizong Shen
May 3, 2021·European Journal of Medicinal Chemistry·Meizhu WangWen Zhou

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Methods Mentioned

BETA
FCS
flow cytometry
glycosylation
confocal microscopy

Software Mentioned

Nikon NIS - elements
GraphPad prism
MitoProtII
Predator

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