ASPN and GJB2 Are Implicated in the Mechanisms of Invasion of Ductal Breast Carcinomas

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.

[1]  P. Taylor,et al.  A prospective study of the development of breast cancer in 16,692 women with benign breast disease. , 1988, American journal of epidemiology.

[2]  J. Haerting,et al.  Gene-expression signatures in breast cancer. , 2003, The New England journal of medicine.

[3]  B. Orr,et al.  Identification of stromally expressed molecules in the prostate by tag-profiling of cancer-associated fibroblasts, normal fibroblasts and fetal prostate , 2011, Oncogene.

[4]  Tina Hernandez-Boussard,et al.  Determination of Stromal Signatures in Breast Carcinoma , 2005, PLoS biology.

[5]  G. M. Nagaraja,et al.  Gene expression signatures and biomarkers of noninvasive and invasive breast cancer cells: comprehensive profiles by representational difference analysis, microarrays and proteomics , 2006, Oncogene.

[6]  Qing Shao,et al.  Connexins act as tumor suppressors in three-dimensional mammary cell organoids by regulating differentiation and angiogenesis. , 2006, Cancer research.

[7]  M. Vijver,et al.  Ductal Carcinoma In Situ of the Breast: Histological Classification and Genetic Alterations , 1998 .

[8]  K. Hirakawa,et al.  Clinical significance of the expression of connexin26 in colorectal cancer , 2010, Journal of experimental & clinical cancer research : CR.

[9]  Y. Benjamini,et al.  Controlling the false discovery rate in behavior genetics research , 2001, Behavioural Brain Research.

[10]  Peter H Watson,et al.  Reduced expression of the small leucine-rich proteoglycans, lumican, and decorin is associated with poor outcome in node-negative invasive breast cancer. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[11]  M. J. van de Vijver.  Ductal carcinoma in situ of the breast: histological classification and genetic alterations. , 1998, Recent results in cancer research. Fortschritte der Krebsforschung. Progres dans les recherches sur le cancer.

[12]  G. Turashvili,et al.  Novel markers for differentiation of lobular and ductal invasive breast carcinomas by laser microdissection and microarray analysis , 2007, BMC Cancer.

[13]  Thangarajan Rajkumar,et al.  Identification and validation of genes involved in gastric tumorigenesis , 2010, Cancer Cell International.

[14]  K. Miyazono,et al.  Mechanisms for Asporin Function and Regulation in Articular Cartilage* , 2007, Journal of Biological Chemistry.

[15]  S. Duffy,et al.  Molecular subtyping of DCIS: heterogeneity of breast cancer reflected in pre-invasive disease , 2010, British Journal of Cancer.

[16]  I. Petersen,et al.  Immunoprofiles of 11 biomarkers using tissue microarrays identify prognostic subgroups in colorectal cancer. , 2005, Neoplasia.

[17]  Jeffrey W Pollard,et al.  Progression to malignancy in the polyoma middle T oncoprotein mouse breast cancer model provides a reliable model for human diseases. , 2003, The American journal of pathology.

[18]  M. J. van de Vijver,et al.  Classification of ductal carcinoma in situ by gene expression profiling , 2006, Breast Cancer Research.

[19]  P. V. van Diest,et al.  Ductal invasive G2 and G3 carcinomas of the breast are the end stages of at least two different lines of genetic evolution , 2001, The Journal of pathology.

[20]  H. Brentani,et al.  Evidence that molecular changes in cells occur before morphological alterations during the progression of breast ductal carcinoma , 2008, Breast Cancer Research.

[21]  Gordon K Smyth,et al.  Linear Models and Empirical Bayes Methods for Assessing Differential Expression in Microarray Experiments , 2004, Statistical applications in genetics and molecular biology.

[22]  Xiao-Jun Ma,et al.  Gene expression profiling of the tumor microenvironment during breast cancer progression , 2009, Breast Cancer Research.

[23]  Rafael A Irizarry,et al.  Exploration, normalization, and genotype calls of high-density oligonucleotide SNP array data. , 2006, Biostatistics.

[24]  Rameen Beroukhim,et al.  Molecular characterization of the tumor microenvironment in breast cancer. , 2004, Cancer cell.

[25]  R. Salunga,et al.  Gene expression profiles of human breast cancer progression , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Jun Yao,et al.  Regulation of in situ to invasive breast carcinoma transition. , 2008, Cancer cell.

[27]  R. Visse,et al.  This Review Is Part of a Thematic Series on Matrix Metalloproteinases, Which Includes the following Articles: Matrix Metalloproteinase Inhibition after Myocardial Infarction: a New Approach to Prevent Heart Failure? Matrix Metalloproteinases in Vascular Remodeling and Atherogenesis: the Good, the Ba , 2022 .

[28]  A. Nobel,et al.  The molecular portraits of breast tumors are conserved across microarray platforms , 2006, BMC Genomics.

[29]  M. Sułkowska,et al.  Increased expression of connexins 26 and 43 in lymph node metastases of breast cancer , 2006, Journal of Clinical Pathology.

[30]  Kay Nieselt,et al.  Progression-specific genes identified by expression profiling of matched ductal carcinomas in situ and invasive breast tumors, combining laser capture microdissection and oligonucleotide microarray analysis. , 2006, Cancer research.

[31]  M. Fassan,et al.  An antimetastatic role for decorin in breast cancer. , 2008, The American journal of pathology.