Determination of DNA synthesis, estrogen receptors, and carcinoembryonic antigen in isolated cellular subpopulations of human breast cancer.

Primary breast adenocarcinomas obtained from ten patients were enzymatically digested using collagenase (1 mg/ml), hyaluronidase (1 mg/ml), elastase (0.1 mg/ml) and DNAse (0.2 mg/ml). The tumor cells were labeled with 3H-thymidine and, in some cases, with 3H-estradiol. The isolated cells were submitted successively to a Ficoll-Hypaque and a bovine serum albumin gradient, from which 12 fractions were obtained. In each fraction, several characteristics were determined: carcinoembryonic antigen (CEA), thymidine (dThd) incorporation, and estrogen receptors (ER). Three main cellular subpopulations were characterized: An intermediate density subpopulation (1.046-1.054 g/ml), in which the proliferating cells are concentrated. In this subpopulation a small number of CEA-positive cells are present, but ER containing cells are virtually absent. A high-density, small cell subpopulation that concentrates most of the ER-containing cells. This subpopulation lacks proliferating cells, but CEA-containing cells are abundant. A low-density subpopulation, lacking proliferating cells and with scarce ER-positive cells, although CEA-positive cells are frequent. These findings strongly suggest that proliferating cells lack ER.

DNA synthesis in estrogen receptor-positive human breast cancer takes place preferentially in estrogen receptor-negative cells.

The primary breast tumors of 27 patients were analyzed for the expression of estrogen receptors (ER) and DNA synthesis. Seventeen tumors were ER-positive, and the simultaneous expression of ER and DNA synthesis could be analyzed in 14 ER-positive tumors. DNA synthesis was measured through the thymidine labeling index (TLI). ER expression was detected by immunohistochemistry with monoclonal antibodies. In these tumors, 38.6% +/- 13.1% of the cells were ER-positive (average TLI = 0.60% +/- 0.70%), as opposed to the presence of 61.4% +/- 13.1% of ER-negative cells (average TLI = 0.65% +/- 0.53%). In 12 of 14 tumors, both ER-positive and ER-negative cells were found to be engaged in DNA synthesis, whereas in two tumors only ER-negative cells were synthesizing DNA. On the basis of the TLI and the proportion of ER-negative and ER-positive cells in the total population, it is suggested that the ER-positive and ER-negative compartments are interrelated in most tumors. In five tumors, the ER-negative compartment would be a precursor of the ER-positive segment, whereas in six tumors the ER-positive segment appears to be a precursor of the ER-negative one. In three tumors, no evidence of an interrelationship between both segments could be found. In the 14 tumors analyzed, it also was found that 69.1% +/- 21.3% of the DNA-synthesizing cells were ER-negative; this probably accounts for the temporary remissions observed after hormonal treatment in breast cancer.