title: iTRAQ-based proteomic profiling of breast cancer cell response to doxorubicin and TRAIL.
abstract: Breast cancer is a molecularly heterogeneous disease, and predicting response to chemotherapy remains a major clinical challenge. To minimize adverse side-effects or cumulative toxicity in patients unlikely to benefit from treatment, biomarkers indicating treatment efficacy are critically needed. iTRAQ labeling coupled with multidimensional LC-MS/MS of the enriched mitochondria and endoplasmic reticulum fraction, key organelles regulating apoptosis, has led to the discovery of several differentially abundant proteins in breast cancer cells treated with the chemotherapeutic agent doxorubicin followed by the death receptor ligand, TRAIL, among 571 and 801 unique proteins identified in ZR-75-1 and MDA-MB-231 breast cancer cell lines, respectively. The differentially abundant proteins represent diverse biological processes associated with cellular assembly and organization, molecular transport, oxidative stress, cell motility, cell death, and cancer. Despite many differences in molecular phenotype between the two breast cancer cell lines, a comparison of their subproteomes following drug treatment revealed three proteins displaying common regulation: PPIB, AHNAK, and SLC1A5. Changes in these proteins, detected by iTRAQ, were confirmed by immunofluorescence, visualized by confocal microscopy. These novel potential biomarkers may have clinical utility for assessing response to cancer treatment and may provide insight into new therapeutic targets for breast cancer.
authors.affiliation: Kolling Institute of Medical Research, University of Sydney, Royal North Shore Hospital, St. Leonards, NSW 2065, Australia.
authors.forename: Sharon
authors.initials: S
authors.lastname: Leong
authors.forename: Andrea C
authors.initials: AC
authors.lastname: Nunez
authors.forename: Mike Z
authors.initials: MZ
authors.lastname: Lin
authors.forename: Ben
authors.initials: B
authors.lastname: Crossett
authors.forename: Richard I
authors.initials: RI
authors.lastname: Christopherson
authors.forename: Robert C
authors.initials: RC
authors.lastname: Baxter
mesh_terms: D026962:Amino Acid Transport System ASC / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000378:metabolism; D045744:Cell Line, Tumor / Q000187:drug effects; D002465:Cell Movement; D017024:Chemotherapy, Adjuvant; D021983:Cyclophilins / Q000378:metabolism; D004317:Doxorubicin / Q000494:pharmacology*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005260:Female; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D053858:Metabolic Networks and Pathways; D015778:Minor Histocompatibility Antigens; D008928:Mitochondria / Q000378:metabolism; D020360:Neoadjuvant Therapy; D009363:Neoplasm Proteins / Q000378:metabolism; D018384:Oxidative Stress; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000494:pharmacology*; D053719:Tandem Mass Spectrometry
chemical_list: C075176:AHNAK protein, human; D026962:Amino Acid Transport System ASC; D000970:Antineoplastic Agents; D008565:Membrane Proteins; D015778:Minor Histocompatibility Antigens; D009363:Neoplasm Proteins; D020543:Proteome; C420506:SLC1A5 protein, human; D053221:TNF-Related Apoptosis-Inducing Ligand; C081786:cyclophilin B; D004317:Doxorubicin; D021983:Cyclophilins