Cancer Epidemiology Biomarkers & Prevention, Vol 7, Issue 11 1027-1033, Copyright © 1998 by American Association for Cancer Research

ARTICLES

Single cell multiple biomarker analysis in archival breast fine-needle aspiration specimens: quantitative fluorescence image analysis of DNA content, p53, and G-actin as breast cancer biomarkers

JY Rao, SK Apple, GP Hemstreet, Y Jin and RK Nieberg
Department of Pathology and Laboratory Medicine, Center of Health Sciences, University of California at Los Angeles, 90024, USA.

Fine-needle aspiration (FNA) is a sensitive and cost-effective method for evaluating breast lesions. However, the diagnosis of early premalignant lesions is less reliable by FNA because of a lack of distinctive cytological features. Accurately defining the risk of such lesions at the individual level may have significant impact in breast cancer prevention and management. The main objective of this preliminary study was to develop a method to study multiple biomarkers on archival FNA slides using quantitative fluorescence image analysis (QFIA). Biomarkers p53, G-actin, and DNA content were labeled with an immunofluorescence technique and measured by QFIA simultaneously on a single cell basis. QFIA allows the labeling and measurement procedures to be carried out in situ, without the need to remove cells from the slide while preserving the morphology of the cells. FNA slides from 72 incident patients were obtained for this study. Fifty-six cases had an adequate number of cells for the actual analysis (25 benign breast lesions, 14 proliferative breast diseases with nuclear atypia, and 17 malignant lesions). The DNA content (> or = 5c) and G-actin (average gray mean, > 90) were positive in 81% and 88% of malignant lesions, respectively. These were significantly higher than the corresponding positive rates in benign lesions (7% and 15%, respectively; P <0.01 for both). None of the benign cases were positive for G-actin and DNA simultaneously, and none of the malignant cases were negative for G-actin and DNA together. p53 was positive in 33% of malignant lesions and 8% of benign lesions (P >0.05). Our study demonstrates the feasibility of evaluating multiple biomarkers by QFIA on archival FNA-fixed specimens. The G-actin and DNA content assayed by QFIA may be potential intermediate end point markers for breast cancer individual risk assessment.

This article has been cited by other articles:

				Y. Jin, K. K. Iwata, A. Belldegrun, R. Figlin, A. Pantuck, Z.-F. Zhang, R. Lieberman, and J. Rao Effect of an epidermal growth factor receptor tyrosine kinase inhibitor on actin remodeling in an in vitro bladder cancer carcinogenesis model. Mol. Cancer Ther., July 1, 2006; 5(7): 1754 - 1763. [Abstract] [Full Text] [PDF]

				M. Hidalgo, M. L. Amador, A. Jimeno, H. Mezzadra, P. Patel, A. Chan, M. E. Nielsen, A. Maitra, and S. Altiok Assessment of gefitinib- and CI-1040-mediated changes in epidermal growth factor receptor signaling in HuCCT-1 human cholangiocarcinoma by serial fine needle aspiration. Mol. Cancer Ther., July 1, 2006; 5(7): 1895 - 1903. [Abstract] [Full Text] [PDF]

				G. P. Hemstreet III, S. Yin, Z. Ma, R. B. Bonner, W. Bi, J. Y. Rao, M. Zang, Q. Zheng, B. Bane, N. Asal, et al. Biomarker Risk Assessment and Bladder Cancer Detection in a Cohort Exposed to Benzidine J Natl Cancer Inst, March 21, 2001; 93(6): 427 - 436. [Abstract] [Full Text] [PDF]

				J. Rao, S. K. Apple, Y. Jin, S. Lin, R. K. Nieberg, and S. L. Hirtschowitz Comparative Polymerase Chain Reaction Analysis of c-myc Amplificationon Archival Breast Fine-Needle Aspiration Materials Cancer Epidemiol. Biomarkers Prev., February 1, 2000; 9(2): 175 - 179. [Abstract] [Full Text]