title: Proteomic profiling of cancer stem cells derived from primary tumors of HER2/Neu transgenic mice.
abstract: Human epidermal growth factor receptor 2 (HER2) overexpression leads to mammary tumorigenesis and its elevated levels lead to increase in cancer stem cells (CSCs), invasion, and metastasis. CSCs are resistant to radiation/chemotherapeutic drugs and are believed to be responsible for recurrence/relapse of cancer. CSCs are isolated using flow cytometry based sorting, although reliable, this technology hinders the convenient identification of molecular targets of CSCs. Therefore to understand the molecular players of increased CSC through HER2 overexpression and to develop meaningful targets for combination therapy, we isolated and characterized breast CSCs through convenient tumorsphere culture. We identified the altered protein expression in CSC as compared to non-CSC using LC-MS/MS and confirmed those results using qRT-PCR and Western blotting. Ferritin heavy chain 1 (FTH1) was identified as a candidate gene, which is involved in iron metabolism and iron depletion significantly decreased the self-renewal of CSCs. We further performed in silico analysis of altered genes in tumorsphere and identified a set of genes (PTMA, S100A4, S100A6, TNXRD1, COX-1, COX-2, KRT14, and FTH1), representing possible molecular targets, which in combination showed a promise to be used as prognostic markers for breast cancer.
authors.affiliation: Department of Biological Science, Centre for Colon Cancer, University of South Carolina, Columbia, SC 29205, USA.
authors.forename: Deepak
authors.initials: D
authors.lastname: Kanojia
authors.forename: Weidong
authors.initials: W
authors.lastname: Zhou
authors.forename: Jiajia
authors.initials: J
authors.lastname: Zhang
authors.forename: Chunfa
authors.initials: C
authors.lastname: Jie
authors.forename: Pang-Kuo
authors.initials: PK
authors.lastname: Lo
authors.forename: Qian
authors.initials: Q
authors.lastname: Wang
authors.forename: Hexin
authors.initials: H
authors.lastname: Chen
mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor / Q000378:metabolism; D015153:Blotting, Western; D018929:Cell Culture Techniques; D003198:Computer Simulation; D003676:Deferoxamine; D005260:Female; D005293:Ferritins / Q000378:metabolism; D006801:Humans; D008325:Mammary Neoplasms, Experimental / Q000201:enzymology / Q000235:genetics / Q000473:pathology*; D051379:Mice; D008822:Mice, Transgenic; D014411:Neoplastic Stem Cells / Q000737:chemistry* / Q000378:metabolism*; D010088:Oxidoreductases; D011379:Prognosis; D020543:Proteome / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics; D060888:Real-Time Polymerase Chain Reaction; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics*; D018874:Spheroids, Cellular; D014407:Tumor Cells, Cultured
chemical_list: D014408:Biomarkers, Tumor; D003676:Deferoxamine; D005293:Ferritins; D010088:Oxidoreductases; D020543:Proteome; D018719:Erb-b2 Receptor Tyrosine Kinases; C454461:FTH1 protein, human