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Relationships of ESR1 and XBP1 expression in human breast carcinoma and stromal cells isolated by laser capture microdissection compared to intact breast cancer tissue

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Abstract

Results from investigations of human genomics which utilize intact tissue biopsy specimens maybe compromised due to a host of uncontrolled variables including cellular heterogeneity of a sample collected under diverse conditions, then processed and stored using different protocols. To determine the cellular origin and assess relationships of mRNA expression of two genes reported to be co-expressed in human breast carcinoma (estrogen receptor-α, ESR1 and X-box binding protein 1, XBP1), gene expression analyses were performed with intact tissue sections and compared with those of laser capture microdissection (LCM)-procured carcinoma and stromal cells from serial sections of the same tissue. Frozen sections of human breast carcinomas were first evaluated for structural integrity and pathology after hematoxylin and eosin (H&E) staining. Total RNA preparations from intact tissue sections and LCM-procured carcinoma and stromal cells were reverse transcribed for measurements of ESR1 and XBP1 expression by quantitative PCR (qPCR). These results were compared with those obtained from microarray analyses of LCM-procured carcinoma cells. Levels of ESR1 and XBP1 were detected in the intact breast cancer tissue sections suggesting coordinate gene expression. Although coordinate expression of these genes was observed in the LCM-procured carcinoma cells, it was not discerned in LCM-procured stromal cells. The origin of coordinate expression of ESR1 and XBP1 observed in whole tissue sections of human breast cancer biopsies is due principally to their co-expression in carcinoma cells and not in the surrounding stromal cells as substantiated using LCM-procured cells. Collectively, a microgenomic process was established from human tissue preparation to RNA characterization and analysis to identify molecular signatures of specific cell types predicting clinical behavior.

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Acknowledgments

The authors wish to acknowledge Dr. D. Alan Kerr II for providing technical advice and Dr. Irina A. Smolenkova for technical assistance in the conduct of this study. Supported in part by grants from the Phi Beta Psi Sorority Charity Trust, the University of Louisville, Office of the Executive Vice President for Research and a CTSP Award from the Commonwealth of Kentucky. SAA was a recipient of a Graduate Fellowship from the Integrated Programs in Biomedical Sciences, University of Louisville, and an AACR Scholar-in-Training Award funded by Susan G. Komen for the Cure™. The experiments outlined in this manuscript comply with the current laws of the country in which they were performed.

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  1. Hormone Receptor Laboratory, Department of Biochemistry & Molecular Biology, Brown Cancer Center and the Institute for Molecular Diversity & Drug Design, University of Louisville, Health Sciences Center A Bldg.—Room 604, Louisville, KY, 40292, USA

    Sarah A. Andres & James L. Wittliff

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Andres, S.A., Wittliff, J.L. Relationships of ESR1 and XBP1 expression in human breast carcinoma and stromal cells isolated by laser capture microdissection compared to intact breast cancer tissue. Endocrine 40, 212–221 (2011). https://doi.org/10.1007/s12020-011-9522-x

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