title: Optimized immunohistochemistry in combination with image analysis: a reliable alternative to quantitative ELISA determination of uPA and PAI-1 for routine risk group discrimination in breast cancer.
abstract: The determination of the invasion markers urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) has further improved the possibilities for individualized therapy of breast cancer. To date, quantitative measurement by ELISA, that needs large amounts of fresh, frozen material, is the only standardized procedure for diagnostic purposes. Therefore, the aim of this study was the establishment of a reliable alternative method based on immunohistochemistry (IHC) and image analysis requiring only small amounts of fixed tumor tissue. Protein expression of uPA and PAI-1 was analyzed in HOPE-fixed tumor samples using tissue microarrays (TMAs) and semiquantitative image analysis. The results of both methods were significantly correlated and risk assessment showed an overall concordance of 78% (83/107; high- and low-risk) and of 94% (74/79) regarding only high-risk patients. The data demonstrate that optimized IHC in combination with image analysis can provide adequate clinical significance compared to ELISA-derived determination of uPA and PAI-1.
authors: Clinical and Experimental Pathology, Research Center Borstel, Parkallee 3a, 23845 Borstel, Germany. Electronic address: dlang@fz-borstel.de.
authors: D S
authors: DS
authors: Lang
authors: U
authors: U
authors: Heilenkötter
authors: W
authors: W
authors: Schumm
authors: O
authors: O
authors: Behrens
authors: R
authors: R
authors: Simon
authors: E
authors: E
authors: Vollmer
authors: T
authors: T
authors: Goldmann
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000032:analysis*; D001943:Breast Neoplasms / Q000737:chemistry*; D018270:Carcinoma, Ductal, Breast / Q000737:chemistry*; D018275:Carcinoma, Lobular / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007089:Image Enhancement; D007150:Immunohistochemistry; D008875:Middle Aged; D017395:Plasminogen Activator Inhibitor 1 / Q000032:analysis*; D018570:Risk Assessment / Q000379:methods; D046888:Tissue Array Analysis; D014568:Urokinase-Type Plasminogen Activator / Q000032:analysis*
chemical_list: D014408:Biomarkers, Tumor; D017395:Plasminogen Activator Inhibitor 1; D014568:Urokinase-Type Plasminogen Activator
keywords: Breast cancer; ELISA; HOPE fixation; Plasminogen activator inhibitor-1; Semiquantitative image analysis; Urokinase-type plasminogen activator