Clinical Cancer Research, Vol 4, Issue 12 3037-3043, Copyright © 1998 by American Association for Cancer Research

ARTICLES

Detection of cancer cells in peripheral blood of breast cancer patients using reverse transcription-polymerase chain reaction for epidermal growth factor receptor

K Leitzel, B Lieu, E Curley, J Smith, V Chinchilli, W Rychlik and A Lipton
Section of Hematology/Oncology, Pennsylvania State University College of Medicine, Hershey Medical Center, 17033, USA.

The epidermal growth factor receptor (EGFR) has been reported to be expressed in high levels in primary breast cancer by immunohistochemistry. In the present study, a reverse transcription (RT)-PCR assay using EGFR primers was developed and evaluated for the detection of circulating micrometastases in the blood of breast cancer patients. Total RNA was extracted from breast cancer cell lines and from the blood of 23 control individuals and 37 breast cancer patients. After reverse transcription, outer and nested primers for EGFR were used for cDNA amplification. RNA integrity was confirmed with parallel RT-PCR amplification using beta2-microglobulin primers. PCR products were electrophoresed on agarose gels containing ethidium bromide and visualized by UV photography. Southern blotting was used to confirm EGFR specificity. The nested EGFR RT-PCR assay was capable of detecting a lower limit of 100 fg of total RNA from the A431 cell line. EGFR RNA was identified from the blood of 4 of 18 (22%) metastatic breast cancer patients, 0 of 6 locally recurrent breast cancer patients, 0 of 13 adjuvant breast cancer patients, and 0 of 23 controls (P = 0.03, metastatic versus control). The 18 metastatic breast cancer patients all had progressive disease at the time of blood sampling. The identity of the four EGFR-positive bands was confirmed by Southern blotting. The presence of RT-PCR positivity for EGFR was not a treatment-related phenomenon, because three of the four EGFR-positive patients were not receiving treatment at the time of blood collection. RT-PCR for EGFR is a sensitive and specific method for the detection of circulating micrometastases in a proportion of patients with metastatic breast cancer.

This article has been cited by other articles:

				M. Lacroix Significance, detection and markers of disseminated breast cancer cells Endocr. Relat. Cancer, December 1, 2006; 13(4): 1033 - 1067. [Abstract] [Full Text] [PDF]

				P. Gazzaniga, O. Gandini, L. Giuliani, M. Magnanti, A. Gradilone, I. Silvestri, W. Gianni, M. Gallucci, L. Frati, and A. M. Aglianò Detection of Epidermal Growth Factor Receptor mRNA in Peripheral Blood: A New Marker of Circulating Neoplastic Cells in Bladder Cancer Patients Clin. Cancer Res., March 1, 2001; 7(3): 577 - 583. [Abstract] [Full Text]

				N. Masuda, Y. Tamaki, I. Sakita, M. Ooka, T. Ohnishi, M. Kadota, N. Aritake, K. Okubo, and M. Monden Clinical Significance of Micrometastases in Axillary Lymph Nodes Assessed by Reverse Transcription-Polymerase Chain Reaction in Breast Cancer Patients Clin. Cancer Res., November 1, 2000; 6(11): 4176 - 4185. [Abstract] [Full Text] [PDF]

				J.-J. Lu, Y. Kakehi, T. Takahashi, X.-X. Wu, T. Yuasa, T. Yoshiki, Y. Okada, T. Terachi, and O. Ogawa Detection of Circulating Cancer Cells by Reverse Transcription-Polymerase Chain Reaction for Uroplakin II in Peripheral Blood of Patients with Urothelial Cancer Clin. Cancer Res., August 1, 2000; 6(8): 3166 - 3171. [Abstract] [Full Text]

				Antonella De Luca, S. Pignata, A. Casamassimi, A. D’Antonio, C. Gridelli, A. Rossi, F. Cremona, V. Parisi, Andrea De Matteis, and N. Normanno Detection of Circulating Tumor Cells in Carcinoma Patients by a Novel Epidermal Growth Factor Receptor Reverse Transcription-PCR Assay Clin. Cancer Res., April 1, 2000; 6(4): 1439 - 1444. [Abstract] [Full Text]