Proteomic Analysis of Formalin-Fixed Paraffin-Embedded (FFPE) Her2 Positive Breast Cancer Tissue Using Differential Mass Spectrometry

摘要

Laser micro-dissection from 30000 cells, followed by Liquid Tissue~R reagent extraction effectively solubilized proteins from FFPE tissue. The sample processing is compatible with LC-MS mass spectrometry. Known breast cancer marker Her2 was measured by SRM quantitation and identified with open dMS discovery platform. A correlation between Her2 levels, FISH scores and IHC scores for selected well characterized cases of Her 2 positive breast cancer was demonstrated. dMS discovery platform revealed 147 proteins that can discriminate between normal and stage 3+ breast cancer. Caldesmon, calnexin, calponin, calreticulin, ezrin and vimentin were selected for further Immunohistochemical (IHC) validation.