title: Automated Quantitative Analysis of p53, Cyclin D1, Ki67 and pERK Expression in Breast Carcinoma Does Not Differ from Expert Pathologist Scoring and Correlates with Clinico-Pathological Characteristics.
abstract: There is critical need for improved biomarker assessment platforms which integrate traditional pathological parameters (TNM stage, grade and ER/PR/HER2 status) with molecular profiling, to better define prognostic subgroups or systemic treatment response. One roadblock is the lack of semi-quantitative methods which reliably measure biomarker expression. Our study assesses reliability of automated immunohistochemistry (IHC) scoring compared to manual scoring of five selected biomarkers in a tissue microarray (TMA) of 63 human breast cancer cases, and correlates these markers with clinico-pathological data. TMA slides were scanned into an Ariol Imaging System, and histologic (H) scores (% positive tumor area x staining intensity 0-3) were calculated using trained algorithms. H scores for all five biomarkers concurred with pathologists' scores, based on Pearson correlation coefficients (0.80-0.90) for continuous data and Kappa statistics (0.55-0.92) for positive vs. negative stain. Using continuous data, significant association of pERK expression with absence of LVI (p = 0.005) and lymph node negativity (p = 0.002) was observed. p53 over-expression, characteristic of dysfunctional p53 in cancer, and Ki67 were associated with high grade (p = 0.032 and 0.0007, respectively). Cyclin D1 correlated inversely with ER/PR/HER2-ve (triple negative) tumors (p = 0.0002). Thus automated quantitation of immunostaining concurs with pathologists' scoring, and provides meaningful associations with clinico-pathological data.
authors.affiliation: Division of Cancer Biology and Genetics, Cancer Research Institute, Queen's University, Kingston K7L 3N6, Canada. elliottb@queensu.ca.
authors.forename: Jamaica D
authors.initials: JD
authors.lastname: Cass
authors.forename: Sonal
authors.initials: S
authors.lastname: Varma
authors.forename: Andrew G
authors.initials: AG
authors.lastname: Day
authors.forename: Waheed
authors.initials: W
authors.lastname: Sangrar
authors.forename: Ashish B
authors.initials: AB
authors.lastname: Rajput
authors.forename: Leda H
authors.initials: LH
authors.lastname: Raptis
authors.forename: Jeremy
authors.initials: J
authors.lastname: Squire
authors.forename: Yolanda
authors.initials: Y
authors.lastname: Madarnas
authors.forename: Sandip K
authors.initials: SK
authors.lastname: Sengupta
authors.forename: Bruce E
authors.initials: BE
authors.lastname: Elliott