title: Centrosome amplification induces high grade features and is prognostic of worse outcomes in breast cancer.
abstract: BACKGROUND
  Centrosome amplification (CA) has been reported in nearly all types of human cancer and is associated with deleterious clinical factors such as higher grade and stage. However, previous reports have not shown how CA affects cellular differentiation and clinical outcomes in breast cancer.
  
  
  METHODS
  We analyzed centrosomes by immunofluorescence and compared to ploidy and chromosomal instability (CIN) as assessed by 6-chromosome FISH in a cohort of 362 breast cancers with median clinical follow-up of 8.4 years. Centrosomes were recognized by immunofluorescence using antibodies for pericentriolar material (PCM; pericentrin) and centrioles (polyglutamylated tubulin). CA was experimentally induced in cell culture by overexpression of polo-like kinase 4 (PLK4).
  
  
  RESULTS
  CA is associated with reduced all-cause and breast cancer-specific overall survival and recurrence-free survival. CA correlates strongly with high-risk subtypes (e.g. triple negative) and higher stage and grade, and the prognostic nature of CA can be explained largely by these factors. A strong correlation between CA and high tumor ploidy demonstrates that chromosome and centrosome doubling often occur in concert. CA is proposed to be a method of inducing CIN via aberrant mitotic cell divisions; consonant with this, we observed a strong correlation between CA and CIN in breast cancers. However, some CA tumors had low levels of CIN, indicating that protective mechanisms are at play, such as centrosome clustering during mitosis. Intriguingly, some high-risk tumors have more acentriolar centrosomes, suggesting PCM fragmentation as another mechanism of CA. In vitro induction of CA in two non-transformed human cell lines (MCF10A and RPE) demonstrated that CA induces a de-differentiated cellular state and features of high-grade malignancy, supporting the idea that CA intrinsically causes high-grade tumors.
  
  
  CONCLUSIONS
  CA is associated with deleterious clinical factors and outcomes in breast cancer. Cell doubling events are the most prevalent causes of CA in cancer, although PCM fragmentation may be a secondary cause. CA promotes high-risk breast cancer in part by inducing high-grade features. These findings highlight the importance of centrosome aberrations in the biology of human breast cancer.
authors.affiliation: Division of Hematology/Oncology, Medical Scientist Training Program and the Department of Medicine, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA. rdenu@wisc.edu.
authors.forename: Ryan A
authors.initials: RA
authors.lastname: Denu
authors.affiliation: Molecular and Cellular Pharmacology Graduate Training Program and the Department of Cell and Regenerative Biology, University of Wisconsin, Madison, WI, USA. lzasadil@wisc.edu.
authors.forename: Lauren M
authors.initials: LM
authors.lastname: Zasadil
authors.affiliation: Wisconsin State Laboratory of Hygiene, University of Wisconsin, Madison, Wisconsin, 53706, USA. craig.kanugh@slh.wisc.edu.
authors.forename: Craig
authors.initials: C
authors.lastname: Kanugh
authors.affiliation: Wisconsin State Laboratory of Hygiene, University of Wisconsin, Madison, Wisconsin, 53706, USA. jjlaffin@wisc.edu.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Laffin
authors.affiliation: Department of Cell and Regenerative Biology and University of Wisconsin Carbone Cancer Center University of Wisconsin, Madison, WI, USA. baweaver@wisc.edu.
authors.forename: Beth A
authors.initials: BA
authors.lastname: Weaver
authors.affiliation: Department of Medicine, Division of Hematology/Oncology and University of Wisconsin Carbone Cancer Center, University of Wisconsin-Madison, 6059 WIMR, 1111 Highland Avenue, Madison, WI, 53705, USA. meburkard@medicine.wisc.edu.
authors.forename: Mark E
authors.initials: ME
authors.lastname: Burkard
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000941:Antigens / Q000235:genetics; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology; D054337:Cell Dedifferentiation / Q000235:genetics; D045744:Cell Line, Tumor; D018385:Centrosome; D002681:China; D005260:Female; D005784:Gene Amplification; D006801:Humans; D017403:In Situ Hybridization; D008875:Middle Aged; D008938:Mitosis / Q000235:genetics; D009367:Neoplasm Staging; D011003:Ploidies; D011379:Prognosis; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis / Q000235:genetics; D014404:Tubulin / Q000235:genetics
chemical_list: D000941:Antigens; D014404:Tubulin; C086178:pericentrin; C490219:PLK4 protein, human; D017346:Protein Serine-Threonine Kinases
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