signature=083a9b2eea41d2a97135c0c36b37a806,Molecular profiling of laser-microdissected matched tumor...

Abstract

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Aims: The aim of the present study was to identify human genes that might prove useful in the diagnosis and therapy of primary breast cancer. Methods: 24 matched pairs of invasive ductal breast cancer and corresponding benign breast tissue were investigated by a combination of laser microdissection and gene expression profiling. Differential expression of up-regulated candidate genes was validated by dot blot analysis of cDNA in 50 pairs of matching benign and malignant breast tissue. Cellular expression of candidate genes was further validated by RNA in-situ hybridization, quantitative RT-PCR, and immunohistochemistry using tissue microarray analysis of 272 non-selected breast cancers. Multivariate analysis of factors regarding overall survival (OS) and recurrence-free survival (RFS) was performed. Results: Dot blot analysis reduced the number of candidate genes to a set of 15 genes that showed at least a two-fold over-expression in more than 15 of 50 (30%) tumor/normal pairs. Among others, we selected KPNA2 for further validation. KPNA2 RNA was up-regulated (fold change > 2) in 32% of analyzed tumor/normal pairs. Nuclear protein expression of KPNA2 was significantly associated with shorter overall survival (OS). Testing various multivariate Cox regression models, KPNA2 expression remained a highly significant, independent and adverse risk factor for OS. Conclusions: Gene expression profiling of laser microdissected breast cancer tissue revealed novel genes that may represent potential molecular targets for breast cancer therapy.