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Molecular Quantitation of Breast Cancer Cells in the Peripheral Blood: Correlation With Clinical Stage at Presentation and Disease Course

Abstract : The overall objective of this project is to develop an assay to detect small numbers of breast cancer cells in a patient's peripheral blood. This assay would be used to test the hypothesis that the presence of circulating breast cancer cells would be predictive of stage at presentation or relapse. The first stage of this project has been to optimize a quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) to quantify breast cancer-specific RNA molecules in the peripheral blood. The result of this initial work was to select keratin 19 (K19) RNA as the best target for detecting small numbers of breast cancer cells. We have also selected an internal reference RNA (beta2-microglobulin), and we have optimized our protocol for extracting RNA from patient blood samples. The second phase of this project has been to develop methods to enrich blood samples for tumor cells, thereby increasing the sensitivity of the assay. Having optimized these parameters, we plan to use the qRT-PCR assay for K19 RNA to study patients with breast cancer, both at the time of presentation and after treatment.