1. Institut de Recerca, Hospital de la Santa Creu i Sant Pau, Barcelona, Spain
2. Department of Pathology. Hospital de la Santa Creu i Sant Pau,
3. Department of Oncology. Hospital de la Santa Creu i Sant Pau,
4. Research Unit. Hospital General Universitario de Alicante
5. Universitat Autònoma de Barcelona, Barcelona, Spain.
* Current address: Department of Molecular Pathology, Vall d'Hebron Research Institute, Barcelona
The mechanism of progression from ductal carcinoma in situ (DCIS) to invasive ductal carcinoma (IDC) remains largely unknown. We compared gene expression in tumors with simultaneous DCIS and IDC to decipher how diverse proteins participate in the local invasive process.
Twenty frozen tumor specimens with concurrent, but separated, DCIS and IDC were microdissected and evaluated. Total RNA was extracted and microarray analysis was performed using Affymetrix GeneChip® Human Gene 1.0 ST Arrays. Microarray data were validated by quantitative real time reverse transcription-PCR (qRT-PCR) and immunohistochemistry. Controls included seven pure in situ carcinomas, eight fragments from normal breast tissue, and a series of mouse breast carcinomas (MMTV-PyMT).
Fifty-six genes were differentially expressed between DCIS and IDC samples. The genes upregulated in IDC samples, and probably associated with invasion, were related to the epithelial-mesenchymal transition (ASPN, THBS2, FN1, SPARC, and COL11A1), cellular adhesion (GJB2), cell motility and progression (PLAUR, PLAU, BGN, ADAMTS16, and ENPP2), extracellular matrix degradation (MMP11, MMP13, and MMP14), and growth/proliferation (ST6GAL2). qRT-PCR confirmed the expression patterns of ASPN, GJB2, ENPP2, ST6GAL2, and TMBS10. Expression of the ASPN and GJB2 gene products was detected by immunohistochemistry in invasive carcinoma foci. The association of GJB2 protein expression with invasion was confirmed by qRT-PCR in mouse tumors (P < 0.05).
Conclusions: The upregulation of ASPN and GJB2 may play important roles in local invasion of breast ductal carcinomas.
Keywords: breast cancer, in situ ductal carcinoma, invasive ductal carcinoma, epithelial-mesenchymal transition, ASPN, GJB2.