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Molecular Oncology, Markers, Clinical Correlates |
Novel Therapeutic Approaches Section, Oncologia Sperimentale D [A. D. L., A. C., N. N.], Oncologia Medica B [S. P., C. G., A. R.], Anatomia Patologica [A. D.], Chirurgia Oncologica C [F. C., V. P.], and Endocrinologia Oncologica [A. D. M.], ITN-Fondazione Pascale, 80131 Naples, Italy
The
epidermal growth factor receptor (EGFR) is overexpressed in 50–70% of
human primary breast, lung, and colon carcinomas, whereas it is not
usually expressed in hematopoietic cells. We developed a novel reverse
transcription-PCR (RT-PCR)-Southern blot assay for the detection of
circulating, EGFR mRNA-expressing tumor cells in carcinoma patients.
The assay was set up by increasing the amount of cDNA step by step in
the PCR reaction. The highest sensitivity and specificity were found
when using 800 ng of cDNA in the PCR reaction. Peripheral blood samples
from 91 patients with either colon (38), lung (30), or breast (23)
carcinomas and from 38 healthy volunteers were analyzed. EGFR
transcripts were found in 44 of 75 (59%) patients with metastatic
carcinoma and in 4 of 38 (10.5%) healthy donors (P <
0.001; 
2 test). The expression of EGFR, cytokeratin 19,
and carcinoembryonic antigen mRNA in blood samples from patients with
metastatic colon carcinoma was compared. EGFR, cytokeratin 19, and
carcinoembryonic antigen transcripts were found in 8 of 11 (73%), 3 of
11 (27%), and 5 of 11 (45%) patients, respectively. Furthermore, two
of seven (29%) Dukes’ B and five of nine (55%) Dukes’ C colon
carcinoma patients were found to express EGFR mRNA in the peripheral
blood. All patients that expressed EGFR transcripts in the peripheral
blood were found to express the EGFR protein in the corresponding
primary carcinoma, as assessed by immunohistochemistry. These data
suggest that the EGFR assay that we developed is a highly specific and
sensitive technique to detect circulating tumor cells in patients
affected by different carcinoma types.
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