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p53 Mutations in Primary and Metastatic Tumors and Circulating Tumor Cells from Colorectal Carcinoma Patients¹

Department of Surgical Oncology, Chelsea and Westminster Hospital [Z. A. J. K., S. K. J., N. L-M., H. P., R. Q. W., A. T., A. I., T. G. A-M.], and Advanced Biotechnology Centre, Charing Cross Hospital, Imperial College School of Medicine [A. T., A. I.], London SW10 9NH, England

Circulating tumor cells could provide a relatively noninvasive and repeatable source of information about tumor cell genotype that might influence treatment and estimation of prognosis. We developed a technique for identifying p53 mutations in tumor cells isolated from the peripheral venous blood of colorectal cancer patients and compared the prevalence and position of these mutations with multiple solid tumor samples from the same patient. We used immunomagnetic beads to isolate tumor cells, reverse transcriptase-nested polymerase chain amplification of the coding region between exons 4 and 9 within the p53 gene, and automated gene sequencing. Nineteen p53 mutations were detected in solid tumor samples from 19 of 41 colorectal carcinoma patients. An identical p53 mutation was invariably present in all samples from primary and metastatic colorectal tumor biopsies within the same patient. p53 mutations were detected in peripheral blood from 8 of these 19 patients with p53-mutated solid tumors. Where identified, the pattern of mutation in peripheral blood samples was invariably the same as in matching solid tumor samples. A single colorectal carcinoma biopsy provided reliable p53 gene mutational information in colorectal carcinoma. Detection of this p53 mutation in tumor cells from peripheral blood was achieved with an approach based on cell selection for epithelial characteristics, reverse transcription-PCR, and gene sequencing.

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