title: Quantum dot-based immunofluorescent imaging and quantitative detection of TOP2A and prognostic value in triple-negative breast cancer.
abstract: BACKGROUND
  Topoisomerase 2 alpha (TOP2A) is a key enzyme in DNA replication and a target of various cytotoxic agents including anthracyclines. Previous studies evaluating the predictive and prognostic values of TOP2A in breast cancer are contradictory, likely secondary to the use of both different detection methods and different cutoff thresholds for positive status. Our own studies have previously confirmed the advantages of quantum dot-based nanotechnology for quantitative analysis of biomarkers relative to conventional immunohistochemistry (IHC). This study was designed to 1) assess the expression of TOP2A, 2) investigate the relationship between TOP2A expression and major clinical pathological parameters, and 3) evaluate the prognostic value of TOP2A by quantum dot-based immunofluorescent imaging and quantitative analytical system (QD-IIQAS) in triple-negative breast cancer (TNBC).
  
  
  METHODS
  TOP2A expression in 145 TNBC specimens was detected using IHC and QD-IIQAS, and a comparative analysis of the two methods was conducted, including an exploration of the relationship between TOP2A expression and major clinical pathological parameters in TNBC. The prognostic value of TOP2A in TNBC was assessed.
  
  
  RESULTS
  A similar antigen localization, a high correlation of staining rates (r=0.79), and a high agreement of measurements (κ=0.763) of TOP2A expression in TNBC were found by QD-IIQAS and conventional IHC (cutoff: 45.0 and 0.45, respectively). TOP2A was significantly higher in larger tumors (P=0.002), higher grade tumors (P=0.005), and lymph node positive patients (P<0.001). The 5-year disease-free survival (5-DFS) of the high and low TOP2A subgroups was significantly different for both QD-IIQAS and IHC (P<0.001, log-rank test for both). TOP2A expression was an independent predictor of survival in TNBC (P=0.001).
  
  
  CONCLUSIONS
  QD-IIQAS was an easy and accurate method for detecting and assessing TOP2A. The TOP2A expression was an independent prognostic indicator of 5-DFS in TNBC. Our study provides a good foundation for future studies exploring the relationship between TOP2A expression and response to anthracyclines.
authors: Department of Breast Surgery, Hubei Cancer Hospital.
authors: Hongmei
authors: H
authors: Zheng
authors: Department of Breast Surgery, Hubei Cancer Hospital.
authors: Xiang
authors: X
authors: Li
authors: Department of Breast and Thyroid Surgery, Renmin Hospital of Wuhan University.
authors: Chuang
authors: C
authors: Chen
authors: Department of Head and Neck Surgery, Hubei Cancer Hospital.
authors: Jian
authors: J
authors: Chen
authors: Department of Breast and Thyroid Surgery, Renmin Hospital of Wuhan University.
authors: Jinzhong
authors: J
authors: Sun
authors: Department of Clinical Laboratory, Renmin Hospital of Wuhan University, Wuhan, Hubei, People's Republic of China.
authors: Si
authors: S
authors: Sun
authors: Department of Breast Surgery, Hubei Cancer Hospital.
authors: Liting
authors: L
authors: Jin
authors: Department of Breast and Thyroid Surgery, Renmin Hospital of Wuhan University.
authors: Juanjuan
authors: J
authors: Li
authors: Department of Breast and Thyroid Surgery, Renmin Hospital of Wuhan University.
authors: Shengrong
authors: S
authors: Sun
authors: Department of Breast Surgery, Hubei Cancer Hospital.
authors: Xinhong
authors: X
authors: Wu
mesh_terms: D000328:Adult; D000368:Aged; D000951:Antigens, Neoplasm / Q000378:metabolism*; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D018572:Disease-Free Survival; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D008875:Middle Aged; D057054:Molecular Imaging / Q000379:methods*; D000075223:Poly-ADP-Ribose Binding Proteins; D045663:Quantum Dots / Q000737:chemistry*; D064726:Triple Negative Breast Neoplasms / Q000175:diagnosis / Q000000981:diagnostic imaging* / Q000378:metabolism*
chemical_list: D000951:Antigens, Neoplasm; D004268:DNA-Binding Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; D004250:DNA Topoisomerases, Type II; C000618811:TOP2A protein, human
keywords: TOP2A; disease-free survival; prognosis; quantum dot; triple-negative breast cancer