neotetrazolium and Fibrocystic-Breast-Disease

We here report a method for the characterization of carcinoma cells in cryosections of human breast tissue. Cryosections from 37 biopsies including carcinomas, fibrocystic disease specimens and reduction mammoplasties were incubated in an atmosphere of either pure nitrogen or 99.5% oxygen. The incubation medium contained neotetrazolium, phenazine methosulphate and the colloid stabilizer Polypep 5115. A marked overlap in formation of reaction product was found between carcinomas, benign tissue and normal breast tissue when sections were incubated in pure nitrogen. However, following 5 min of oxygen incubation no reaction was seen in normal breast tissue, whereas carcinoma cells consistently showed reaction. Moreover, a statistically significant difference was seen between the level of reaction in carcinoma cells and that of ductal epithelial cells in fibrocystic disease not containing apocrine metaplasia. Apocrine metaplasia, on the other hand, showed the highest activity recorded at all.

Topics: Breast; Breast Neoplasms; Carcinoma; Carcinoma, Intraductal, Noninfiltrating; Epithelium; Female; Fibrocystic Breast Disease; Glucosephosphate Dehydrogenase; Humans; Oxygen; Tetrazolium Salts

Epithelial cell islets in primary monolayer cultures of human breast biopsies were characterized by combined immuno-, enzyme- and DNA cytochemistry as well as by analysis of attachment-, spread- and growth patterns. For cultivation we used explants from reduction mammoplasties, benign lesions, primary carcinomas and metastases. Milk fat globule membrane antigen (MFGM-A) was detected with a monoclonal antibody, and the tetrazolium reaction for glucose 6-phosphate dehydrogenase (G6PDH) as well as DNA content of the cultured cells were quantified. Spreading and growth of individual islets were studied by image analysis. Fibroblast-like cells did not express MFGM-A, and whereas epithelial (MFGM-A positive) cell islets of normal and benign origin showed cells with no or low G6PDH reaction, respectively, the majority of epithelial cell islets from 11 out of 21 carcinomas showed strong reaction. Cell islets with strong G6PDH reaction were sometimes hyperdiploid. Moreover, whereas cell islets with no or low reaction from both benign lesions and carcinomas readily attached and spread in a serum-free medium and showed population doubling times of 30 to 110 h, cell islets with strong reaction from carcinomas and metastatic lesions required serum for attachment and their growth rate was too low to be determined.

Topics: Antigens, Surface; Breast Neoplasms; Carcinoma; Cell Adhesion; Cell Division; Cells, Cultured; DNA, Neoplasm; Epithelium; Female; Fibrocystic Breast Disease; Glucosephosphate Dehydrogenase; Humans; Lymphatic Metastasis; Membrane Proteins; Mucin-1; Tetrazolium Salts