DNA image cytometry in the differential diagnosis of endocervical adenocarcinoma.

BACKGROUND: The value of DNA image cytometry in the differential diagnosis of endocervical adenocarcinoma was tested on a series of 65 cases of normal endocervical cells (n = 25), inflammatory changes (n = 18), and endocervical adenocarcinoma (n = 22). METHODS: The investigation was performed on gynecologic routine smears by using a television image analysis system MIAMED-DNA (Leica, Wetzlar, Germany), combined with an automated Leica Medilux microscope. First, the Papanicolaou stained specimens were rescreened, and the x/y coordinates of at least 150 endocervical nuclei were stored per case by using a scanning program. After restaining according to Feulgen, the epithelial cells were relocalized and the DNA content, and the nuclear area were determined. The DNA content of 25-30 squamous epithelial cells of intermediate type served as an internal standard for the normal diploid value in each case. Various DNA cytometric parameters and the mean nuclear area were calculated. For statistical analysis, the cases of adenocarcinoma (n = 22) were defined as positive, and the cases with normal endocervical epithelium or inflammatory changes (n = 43) were defined as negative. RESULTS: The presence of nuclei with a DNA content greater than 9c was observed exclusively in adenocarcinoma (sensitivity, 95.9%; specificity, 100%), indicating that this parameter is suited best for the differentiation between malignant and nonmalignant endocervical epithelium. High sensitivity rates at a specificity level of 100% also were calculated for the 2.5cER (95.5%), the mean ploidy (90.9%), 5cER (90.9%), and the diploid deviation quotient (90.9%). For the 2cDI (86.4%), the entropy (81.8%) and the ploidy imbalance (77.3%) lower values were obtained. CONCLUSIONS: DNA single cell cytometry represents a highly relevant tool in the identification of malignant transformation in endocervical lesions that could be used as a complementary diagnostic method in cytologically difficult cases. Investigations on endocervical adenocarcinoma in situ should be performed in the near future.

Rapid and prognostically valid quantification of immunohistochemical reactions by immunohistometry of the most positive tumour focus. A prospective follow-up study on breast cancer using antibodies against MIB-1, PCNA, ER, and PR.

The prognostic significance of immunohistochemical markers for cell proliferation [MIB-1, proliferating cell nuclear antigen (PCNA)] and hormone receptor analysis [oestrogen receptor (ER), progesterone receptor (PR)] was tested by means of immunohistometry in a series of 103 breast cancer patients in comparison with the lymph node status N, the tumour size T, histomorphological grading, and the biochemical ER and PR status. Immunohistochemical reactions were performed on 2 microns sections from paraffin-embedded tissue, using an indirect peroxidase method. The proportion of immunostained tumour cell nuclei was determined using a TV-image analysis system. Measurements were performed using a 20 x objective on 40 viewing fields (1.94 mm2, MIB-1 and PCNA) or 20 viewing fields (0.97 mm2, ER and PR). The mean immunopositivity of all viewing fields and the value of the most immunopositive viewing field (MIB-1max, PCNAmax, PRmax, ERmax) were calculated. The mean values and the maximal values were highly correlated (r = 0.903, P < 0.001). After 1:2:1 quantilization, 84.2 per cent of the 412 single measurements revealed mean and maximal values in the same category (P < 0.0001). For each of the four immunohistochemical markers, the prognostic significance of the maximal values was higher than that of the mean values. The highest prognostic significance was found for MIC-1max (P = 0.0002), followed by PRmax (P = 0.0046), ERmax (P = 0.0154), and PCNAmax (P = 0.0161). From the results of a Cox model, a 'prognostic index (PI)' was developed, ranging from -1 to 8: PI = 2 x N + T + MIB-1max-PRmax. The four groups of patients with PI values of < 2, 2-3, 4-5, and > 5 revealed significantly different 7.5-year survival probabilities (P < 0.0001). The simplicity of the PI makes it a clinically useful, routinely applicable, and understandable parameter in the surgical pathology of breast carcinoma.