The importance of tissue handling of surgically removed breast cancer for an accurate assessment of the Ki-67 index

J Clin Pathol. 2016 Mar;69(3):255-9. doi: 10.1136/jclinpath-2015-203174. Epub 2015 Sep 29.

Abstract

Aim: Insufficient attention for the Ki-67 immunohistochemistry has been given to the importance of tissue handling for surgical breast cancer specimens. We sought to investigate the effect of fixation status on the Ki-67.

Methods: We examined the effect of fixative, time to and duration of fixation using surgical specimens, and finally, compared the paired Ki-67 index in the tumour between core needle and surgical specimen.

Results: The Ki-67 was significantly higher when 10% neutral buffered formalin was used (p=0.0276). Insufficient fixation caused a drastic reduction in the Ki-67 index (p=0.0177), but not significant in oestrogen receptor (ER) and human epidermal growth factor receptor 2 (HER2). Sixteen hours delayed time to fixation also caused a reduction of the Ki-67 (p=0.0284), but not significant in ER. Prolonged fixation significantly led to a gradual reduction in the Ki-67 in a time-dependent manner, but not in both ER and HER2. Finally, cutting the tumour before fixation improved fixation status and consequently caused an increased level of the Ki-67 index (p=0.0181), which resulted in a strong correlation of the Ki-67 between core needle and surgical specimen (r=0.8595).

Conclusions: Tissue handling of surgical specimen is critical for assessing the Ki-67 compared with ER and HER2. We should pay more attention to tissue fixation status for the standard assessment of the Ki-67 index.

Keywords: BREAST CANCER; IMMUNOHISTOCHEMISTRY; PROLIFERATION; TUMOUR BIOLOGY.

Publication types

  • Comparative Study

MeSH terms

  • Biopsy, Large-Core Needle
  • Breast Neoplasms / chemistry*
  • Breast Neoplasms / pathology
  • Breast Neoplasms / surgery*
  • Female
  • Fixatives*
  • Humans
  • Immunohistochemistry*
  • Ki-67 Antigen / analysis*
  • Mastectomy*
  • Predictive Value of Tests
  • Protein Stability
  • Proteolysis
  • Receptor, ErbB-2 / analysis
  • Receptors, Estrogen / analysis
  • Reproducibility of Results
  • Specimen Handling / methods*
  • Time Factors
  • Tissue Fixation / methods*

Substances

  • Fixatives
  • Ki-67 Antigen
  • Receptors, Estrogen
  • ERBB2 protein, human
  • Receptor, ErbB-2