Detection of circulating tumour cells in peripheral blood with an automated scanning fluorescence microscope

• Published in: British journal of cancer Vol. 99; no. 5; pp. 789 - 795
• Main Authors: Ntouroupi, T G, Ashraf, S Q, McGregor, S B, Turney, B W, Seppo, A, Kim, Y, Wang, X, Kilpatrick, M W, Tsipouras, P, Tafas, T, Bodmer, W F
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 02-09-2008; Nature Publishing Group
• Subjects: Antigens; Automation; Biological and medical sciences; Biomedical and Life Sciences; Biomedicine; Biopsy; Cancer Research; Cell Line, Tumor; Cells; Chromosomes; Colorectal Neoplasms - blood; Colorectal Neoplasms - pathology; Drug Resistance; Epidemiology; Experiments; Female; Humans; In Situ Hybridization, Fluorescence; Male; Medical research; Medical sciences; Membrane filters; Microscopy, Fluorescence - methods; Molecular Diagnostics; Molecular Medicine; Neoplastic Cells, Circulating; Nephrology. Urinary tract diseases; Oncology; Ovarian cancer; Ovarian Neoplasms - blood; Ovarian Neoplasms - pathology; Prostate cancer; Prostatic Neoplasms - blood; Prostatic Neoplasms - pathology; Recurrence; Tumors; Tumors of the urinary system; Urinary tract. Prostate gland; Yeast; New Zealand; United Kingdom > UK; United States > US; colorectal, prostate and ovarian cancer; circulating tumour cells; automated fluorescence microscope; Urinary system disease; Prostate disease; Ovary cancer; Fluorescence; Malignant tumor; Female genital diseases; Ovarian diseases; circulating tumour cells; automated fluorescence microscope; colorectal; Blood cell; Scanning microscope; Cancerology; prostate and ovarian cancer; Diagnosis; Male genital diseases; Prostate cancer; Tumor cell; Cancer
• ISSN: 0007-0920; 1532-1827
• DOI: 10.1038/sj.bjc.6604545

We have developed an automated, highly sensitive and specific method for identifying and enumerating circulating tumour cells (CTCs) in the blood. Blood samples from 10 prostate, 25 colorectal and 4 ovarian cancer patients were analysed. Eleven healthy donors and seven men with elevated serum prostate-specific antigen (PSA) levels but no evidence of malignancy served as controls. Spiking experiments with cancer cell lines were performed to estimate recovery yield. Isolation was performed either by density gradient centrifugation or by filtration, and the CTCs were labelled with monoclonal antibodies against cytokeratins 7/8 and either AUA1 (against EpCam) or anti-PSA. The slides were analysed with the Ikoniscope
®
robotic fluorescence microscope imaging system. Spiking experiments showed that less than one epithelial cell per millilitre of blood could be detected, and that fluorescence
in situ
hybridisation (FISH) could identify chromosomal abnormalities in these cells. No positive cells were detected in the 11 healthy control samples. Circulating tumour cells were detected in 23 out of 25 colorectal, 10 out of 10 prostate and 4 out of 4 ovarian cancer patients. Five samples (three colorectal and two ovarian) were analysed by FISH for chromosomes 7 and 8 combined and all had significantly more than four dots per cell. We have demonstrated an Ikoniscope
®
based relatively simple and rapid procedure for the clear-cut identification of CTCs. The method has considerable promise for screening, early detection of recurrence and evaluation of treatment response for a wide variety of carcinomas.