Comparison of Gene Expression in Breast Cancer Using Laser Capture Microdissection and Quantitative RT-PCR

Amplification and deletion are the most common mechanisms leading genes to gene deregulation in breast cancer, but the relevance, the role and timing of most of the described genetic abnormalities are still unclear. The significance and usefulness of those genetic abnormalities will have to be established by the use of reliable and reproducible methodologies. In this study, we used the advanced technique of laser capture microdissection to obtain pure tumor cell populations and normal epithelium in paired breast tissue samples from the same patient, and submitted them to further analysis by real-time quantitative RT-PCR for a rapid, objective and reliable quantification of even small abnormalities. We will also introduce and recommend a simple staining method and a suitable thickness of section for LCM and quality RNA extraction from the tissue samples. We believe that this method will provide more detailed evidence of gene expression in the study of cancers, especially in multi-step cancer carcinogenesis.