p53 gene mutations are not required for early dissemination of cancer cells.

The p53 protein is involved in several central cellular processes, including gene transcription, DNA repair, cell cycling, genomic stability, chromosomal segregation, senescence, and apoptosis. p53 mutations frequently result in an immunocytochemically detectable accumulation of the p53 protein in tumor cells. To evaluate whether p53 gene mutations are required for the onset of hematogeneous tumor cell dissemination, we compared the p53 status of primary and micrometastatic tumor cells. Disseminated carcinoma cells could be detected in bone marrow aspirates obtained from 46 (40%) of 114 patients with various types of epithelial tumors without overt skeleton metastases. There was no correlation between the detection of p53 protein in primary lung carcinomas and the presence of tumor cells in bone marrow. Further analyses revealed that the disseminated carcinoma cells rarely accumulate mutated p53 protein and that 10 cell lines derived thereof did not harbor p53 mutations even in the presence of such mutations in the autologous primary tumors. These observations indicate that tumor cells can leave the primary tumor before mutations of the p53 gene occur and that these mutations are not essential for such early hematogeneous dissemination of cancer cells. Thus, the value of mutated p53 as a target for diagnosis and treatment of micrometastatic disease in cancer patients is questionable.

Methodological analysis of immunocytochemical screening for disseminated epithelial tumor cells in bone marrow.

The emerging clinical relevance of bone marrow micrometastasis has prompted several investigations, using a variety of immunocytochemical approaches. The present study was designed to evaluate some of the variables affecting the immunocytochemical detection of individual epithelial tumor cells in bone marrow. Using an alkaline phosphatase-antialkaline phosphatase staining technique, we evaluated bone marrow aspirates from 358 patients with primary carcinomas of the breast (n = 150), lung (n = 66), prostate (n = 42), or colorectum (n = 100). Individual tumor cells in cytological preparations were detected with monoclonal antibody (MAb) CK2 to the epithelial cytokeratin component 18 (CK18), which has been validated in extensive clinical studies. In addition, the utility of the broad-spectrum MAb A45-B/B3 was explored in this study. The high specificity of MAbs CK2 and A45-B/B3 was supported by analysis of bone marrow from 75 noncarcinoma control patients and by double-marker analysis with MAbs to mesenchymal marker proteins (CD45 and vimentin). In contrast, MAbs E29 and HMFG1, directed to mucin-like epithelial membrane proteins, cross-reacted with hematopoietic cells in 26.7-42.7% of all samples tested. The majority of the 154 positive samples (43.0%) from cancer patients displayed less than 10 CK18-positive cells per 8 x 10(5) marrow cells analyzed. The detection rate, however, was affected by blood contamination of the aspirate, the number of aspirates analyzed, and the number of marrow cells screened per aspiration site. Comparative immunostaining of bone marrow specimens with MAbs CK2 and A45-B/B3 indicated that downregulation of CK18 in micrometastatic carcinoma cells occurs in about 50% of the 172 samples analyzed, regardless of the primary tumor origin.(ABSTRACT TRUNCATED AT 250 WORDS)