Identifying Circulating Breast Cancer Cells in Women With Metastatic Breast Cancer

• Feasibility of novel technique to identify and isolate circulating breast cancer cells [ Time Frame: Baseline ] [ Designated as safety issue: No ]
• Comparison of this novel technique with the CellSearch™ system [ Time Frame: Baseline ] [ Designated as safety issue: No ]

• Feasibility of novel technique to identify and isolate circulating breast cancer cells [ Designated as safety issue: No ]
• Comparison of this novel technique with the CellSearch™ system [ Designated as safety issue: No ]

RATIONALE: Studying samples of blood and pleural or peritoneal fluid from patients with metastatic breast cancer in the laboratory may help doctors identify biomarkers related to breast cancer and learn more about how breast cancer begins and spreads in the body.

PURPOSE: This research study is looking at a new way of identifying circulating breast cancer cells in blood and in pleural or peritoneal fluid in women with metastatic breast cancer.

OBJECTIVES:

Primary

• To compare identification of circulating breast cancer cells (CBCCs) in blood or pleural or peritoneal fluid by a novel technique using stem cell marker retinaldehyde dehydrogenase (ALDH) and surface antigen expression (CD44+, CD24-) to the standard technique using the CellSearch® system in women with metastatic breast cancer.

Secondary

• To determine whether CBCCs have the potential to grow into metastatic lesions.

OUTLINE: Patients undergo sample collection to help develop a new technique using stem cell marker retinaldehyde dehydrogenase (ALDH) and surface antigen expression (CD44+, CD24-) in isolating circulating breast cancer cells (CBCCs) from blood and pleural or peritoneal fluid. Blood may also be drawn to measure the number of circulating tumor cells using the standard CellSearch® system.

Mononuclear cells are isolated by density centrifugation. Cells are stained against surface antigens that provide specific expression patterns for CBCCs (CD44, CD24). Cells are analyzed on a fluorescence activated cell sorting (FACS) Calibur flow cytometer and sequentially gated (ALDHhigh→CD44+ vs CD24-/low or CD44+ vs CD24-/low→ALDHhigh) for detection of CBCCs. For further confirmation of epithelial origin, ALDHhighCD44+CD24-/low cells are isolated using a FACSAria flow sorter, cytocentrifuged onto glass slides then stained for the expression of epithelial-specific cytokeratins 5, 8, 14, 18 and 19 by standard immunohistochemical techniques. Using the phenotype that is found to most highly enrich for epithelial cells, cells are isolated by FACS and assayed for clonogenic growth.

Metastatic breast cancer patients

• Other: flow cytometry
  laboratory analysis
• Other: fluorescence activated cell sorting
  laboratory analysis
• Other: immunohistochemistry staining method
  laboratory analysis
• Other: immunologic technique
  laboratory analysis
• Other: biomarker analysis
  laboratory analysis

DISEASE CHARACTERISTICS:

• Histologically confirmed adenocarcinoma of the breast

  • Metastatic disease (stage IV)
• Hormone receptor status not specified

PATIENT CHARACTERISTICS:

• Female
• Menopausal status not specified

PRIOR CONCURRENT THERAPY:

• Not specified