Gene Transcript Assay by Real-Time Rt-PCR in Epithelial Breast Cancer Cells Selected by Laser Microdissection

Author:

Becette V.1,Vignaud S.2,Régnier C.1,Labroquère M.2,Fourme E.3,Menet E.1,Bièche I.45,Spyratos F.2

Affiliation:

1. Département de Pathologie, INSERM E0017, Centre René Huguenin, Saint-Cloud

2. Laboratoire d'Oncobiologie, INSERM E0017, Centre René Huguenin, Saint-Cloud

3. Département de Statistiques Médicales, INSERM E0017, Centre René Huguenin, Saint-Cloud

4. Laboratoire d'On-cogénétique, INSERM E0017, Centre René Huguenin, Saint-Cloud

5. Laboratoire de Génétique Moléculaire, UPRES EA 3618, Faculté des Sciences Pharmaceutiques et Biologiques de Paris, Paris - France

Abstract

The cell type heterogeneity within clinical cancer tissue samples may affect the accuracy of gene expression analysis. In order to validate our laser microdissection (LMD) method using the Leica AS LMD system (LEICA Microsystems), we compared the mRNA levels of three major genes involved in breast cancer (ERα, PR, HER2), measured by means of real-time quantitative RT-PCR, in 5000 microdissected malignant epithelial cells and in corresponding bulk tumor ho-mogenates from 14 patients. We also compared the mRNA level results to protein expression measured by immunohistochemistry (IHC) on the same tumors. For the three genes, significant correlations were found between mRNA results obtained on microdissected cells and IHC. Comparison between IHC and mRNA results obtained on microdissected cells and bulk tumors showed that in all cases microdissection enhanced the sensitivity of assessing target gene transcript levels and was essential for their accurate evaluation in heterogeneous tumors.

Publisher

SAGE Publications

Subject

Cancer Research,Clinical Biochemistry,Oncology,Pathology and Forensic Medicine

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