An investigation of heat shock protein 27 and P-glycoprotein mediated multi-drug resistance in breast cancer using liquid chromatography-tandem mass spectrometry-based targeted proteomics

摘要: Abstract One missing puzzle piece to study heat shock protein 27 (HSP27) in P-glycoprotein (P-gp) mediated multi-drug resistance (MDR) was the amount of HSP27 and extent its phosphorylation biological context. Liquid chromatography–tandem mass spectrometry (LC/MS/MS)-based targeted proteomics allows researchers monitor associated proteins their modification simultaneously quantitatively. In this study, a assay first developed validated for quantification phosphorylated forms. Using assay, level determined non-tumoral cells MCF-10A, parental drug-sensitive cancer MCF-7/WT drug-resistant MCF-7/ADR. A decrease expression observed P-gp overexpressed MCF-7/ADR cells. quantitative time-course analysis both doxorubicin (DOX)-treated also implied that may participate modulation. Furthermore, stoichiometry site-specific indicated DOX treatment rapidly induced at Ser82. Moreover, conventional analytical methods were performed comparison. Biological significance LC/MS/MS-based turns out be promising approach preclinical clinical environment. Unfortunately, rare studies applied technology detect multiple or one experiment. This demonstrated potential understand cell events more accurate context system. By forms sites Ser15 Ser82, possible role MDR suggested. Further development future provide insight into signal transduction pathways upon perturbation network changes panel proposed biomarkers given disease state.