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

摘要

The overall objective of this project was to develop an assay to detect small numbers of breast cancer cells in a patient's peripheral blood. This assay was used to test the hypothesis that the presence of circulating breast cancer cells is predictive of stage at presentation or relapse. The first stage of this project was 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 (K 19) RNA as the best target for detecting small numbers of breast cancer cells. We also selected an internal reference RNA (Beta2-microglobulin), and we optimized our protocol for extracting RNA from patient blood samples. The second stage of this work was to develop ways to enrich peripheral blood specimens for tumor cells, thereby increasing the sensitivity of the assay. Having optimized these parameters, in the third phase we are using the qRT-PCR assay for K 19 RNA to study patients with breast cancer, both at the time of presentation and after treatment.