Comparison of PCNA/cyclin immunohistochemistry with flow cytometric S-phase fraction in breast cancer.
Kinetic index determined by enumeration of neoplastic cells positive for proliferative cell nuclear antigen (PCNA) in 70 breast carcinomas (avidin-biotin immunoperoxidase technique) was compared to synthesis-phase fraction (S-phase, or SPF) values obtained by flow cytometry (FCM) using a multiparametric, 2 color method (dual-label propidium iodide/cytokeratin-FITC). The percent PCNA positive tumor cells (12.5% mean, range 1-28%) was significantly greater in aneuploid tumors (14.2% mean, N = 35) compared to diploid range tumors (10.7% mean, N = 35) (p less than 0.05), and was correlated with SPF derived from ungated DNA histograms (12.5% mean +/- 5.5%, r = 0.45, p less than 0.001). Marginally stronger statistical correlations were observed between the PCNA index and SPF values calculated from cytokeratin-gated (15.8% mean, r = 0.53, p less than 0.001) DNA histograms or from SPF values obtained following linear baseline debris subtraction (mean = 8.1%, r = 0.48, p less than 0.001). Significant associations were identified between PCNA index and prognostically important clinicopathologic parameters including nuclear grade (p = 0.014), presence of necrosis (p = 0.005), and angiolymphatic invasion (p = 0.003). We conclude: 1) PCNA index is comparable to FCM SPF and correlates with factors of known prognostic importance in carcinoma of the breast; 2) baseline debris and contaminating events derived from non-epithelial cells both represent significant artifacts in proliferative fraction estimates derived from FCM DNA histograms; and 3) multiparametric analysis may represent one means of improving the specificity and clinical value of FCM SPF determinations.
Department of Pathology, Henry Ford Hospital, Detroit, Michigan.
D W
DW
Visscher
S
S
Wykes
J
J
Kubus
J D
JD
Crissman
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000951:Antigens, Neoplasm / Q000032:analysis*; D016477:Artifacts; D001943:Breast Neoplasms / Q000276:immunology*; D002455:Cell Division / Q000276:immunology; D004273:DNA, Neoplasm / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007124:Immunoenzyme Techniques; D008875:Middle Aged; D009687:Nuclear Proteins / Q000032:analysis*; D011379:Prognosis; D018809:Proliferating Cell Nuclear Antigen; D011446:Prospective Studies; D016196:S Phase / Q000235:genetics*
D000951:Antigens, Neoplasm; D004273:DNA, Neoplasm; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen