ABSTRACT
Breast cancer is the rst leading cause of cancer death in women. Release of tumor cells into blood circulation may occur at early stage of the disease and may be responsible for its progression [1]. Metastasis in breast cancer patients leads to cancer-related death because early dissemination of tumor cells usually remains undetected at initial diagnosis on clinical, imaging, and biochemical examination. Thus, detection of circulating tumor cells (CTCs) in a blood sample becomes of potential value despite their rarity (median, ≤1 CTC/mL). CTCs have been successfully detected and isolated from blood in patients with metastatic carcinomas [2,3]. A simple blood test would allow the detection and analysis of CTCs to be frequently repeated. It would also help to noninvasively stage the disease at diagnosis as well as to monitor therapy and long-term patient management [4,5]. Several reports have shown that efcacy of treatment could be measured by the number of CTCs in blood [6-8]. CTCs provide easy access to patients’ cancer cells for performing several molecular analyses. Studies on alterations in CTC oncogenes suggested
21.1 Introduction ...................................................................................................................................375 21.2 Materials ...................................................................................................................................... 377 21.3 Methods ........................................................................................................................................ 377
21.3.1 Glass Surface Silanization ............................................................................................... 377 21.3.2 Physical Characterization of the Silane Films and Quality Control ............................... 377 21.3.3 Physical Characterization of Antibody-Coated Surfaces ................................................ 378 21.3.4 Characteristics of Laminar Flow and Simulation in a Parallel Plate Flow Chamber ..... 378 21.3.5 Isolation of MCF7 Breast Cancer Cells Spiked in Normal Blood Leukocytes .............. 379 21.3.6 Immuno•uorescence Staining and Identication of MCF7 Cells by
Fluorescence Microscopy ................................................................................................ 380 21.4 Results .......................................................................................................................................... 380
21.4.1 Physical Characteristics of the Antibody-Coated AHTS Surface .................................. 380 21.4.2 Simulation of the Fluid Flow in a Single Parallel Plate Laminar Flow Chamber ...........381 21.4.3 Isolation of MCF7 Breast Cancer Cells Spiked in Background
Normal Blood Leukocytes ...............................................................................................381 21.4.4 Identication of MCF7 Breast Cancer Cells Spiked in Normal Blood Leukocytes ....... 383
21.5 Discussion .................................................................................................................................... 383 21.6 Future Trends ............................................................................................................................... 384 Acknowledgments .................................................................................................................................. 384 References .............................................................................................................................................. 385
heterogeneity among cancer cells from a single patient, for example, with respect to heterogeneity of HER-2 and uPA gene copy number and expression in breast cancer [9,10]. Thus, identifying the CTC subpopulations contained in a blood sample could help to identify targets for treatment and exploring mechanisms that underlie metastases and drug sensitivity. Increasing the useful information on CTCs might help to tailor systemic therapies to the individual needs of a cancer patient.
