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We have demonstrated the utility of conducting expression profiling of breast tumour samples using a custom selected set of genes to investigate pathological processes related to subtype.
We then performed immunohistochemical analysis of CtBP1 and CtBP2 expression in 22 breast tumour samples, using formalin-fixed, paraffin-embedded archival samples from the Cellular Pathology Department at Southampton General Hospital.
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Formalin-fixed paraffin-embedded (FFPE) breast tumour samples used in this study were acquired through several participating Spanish hospitals (Gregorio Marañon, Hospital San Pablo, Fundacion Jimenez Diaz, Hospital La Paz, H Santa Caterina (Girona), P de Hierro, H Severo Ochoa, H Ramón y Cajal).
This study demonstrates that it is possible to derive biologically meaningful data from a cohort of archived FFPE tumour samples using the Almac Breast DSA™.
Using all genes in two signatures of ERBB2 and EGFR activation [ 1, 3] to infer pathway activity in a large set of breast tumour samples and using either Spearman or Pearson correlations showed that predicted ERBB2 activity was not highest in the intrinsic HER2+ subtype, and similarly that EGFR activity was not highest in the basal subtype (Additional file 3).
Gene expression analysis of breast tumour samples was performed using custom-made Agilent 44K high-density microarrays according to manufacturer's protocols.
Fluorescence in situ hybridisation on paraffin-embedded breast tumour samples was performed using a modified Paraffin Pretreatment Reagent kit protocol (Vysis) as described previously (Rauta et al, 2006).
Breast tumour samples were inserted as triplicates using a 600 μm needle in 8 Tissue Micro Array (TMA) blocks.
Formalin-fixed paraffin-embedded breast tumour samples were inserted in triplicate using a 600- μm needle (Alphelys, Plaisir, France) in three TMA blocks.
In the present study, normal breast tissue samples (n = 32) and their respective matched breast tumour samples (n = 32) were used for immunohistochmemical analysis.
Importantly, our current study includes AJCC stage 1 IDC only, whereas our previous study used breast tumour samples of AJCC stages 1 3.
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