Real-Time PCR Analysis of Peripheral Blood: A Non-Invasive Technology that can be Used to Assess Tumor Burden in Breast Cancer Patients
Abstract
The molecular detection of circulating tumor cells (CTC) in breast cancer patients has been limited by two factors: 1) low concentration of CTC in peripheral blood, and 2) normal lymphocytes express some cancer-associated genes at detectable levels. In this study, we investigated the ability of a novel bouyant density gradient centrifugation (BDGC) medium (Oncoquick, HEXAL Gentech) to enrich CTC prior to real- time RT-PCR analysis. METHODS: MDA361 breast cancer cells were spiked into normal PBL by micromanipulation, and processed using Ficoll or BDGC medium. Tumor cell enrichment was assessed with flow cytometry, standard cytology, and real-time RT-PCR. RESULTS: BDGC medium dramatically reduced the number of recovered mononuclear cells. in a pilot study, 64% of Stage IV breast cancer patients and 100% of those patients naive to chemotherapy had evidence of overexpression of at least one of five cancer-associated genes in peripheral blood. CONCLUSIONS: The use of bouyant density gradient centrifugation media: 1) results in superior reduction in background expression of cancer-associated gene from lymphocytes, 2) enhances tumor cell detection beyond established limits, and 3) has been successfully applied in Stage IV disease. Given the overall efficacy of Oncoquick, its use in Stage I-II disease is promising.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2002
- Accession Number
- ADA412843
Entities
People
- Michael Mitas
Organizations
- Medical University of South Carolina