Molecular Characterization of Human Breast Tumor Vascular Cells

Abstract

A detailed understanding of the assortment of genes that are expressed in breast tumor vessels is needed to facilitate the development of novel, molecularly targeted anti-angiogenic agents for breast cancer therapies. Rapid immunohistochemistry using factor VIII-related antibodies was performed on sections of frozen human luminal-A breast tumors (n ‫؍‬ 5) and normal breast (n ‫؍‬ 5), followed by laser capture microdissection of vascular cells. RNA was extracted and amplified, and fluorescently labeled cDNA was synthesized and hybridized to 44,000-element longoligonucleotide DNA microarrays. Statistical analysis of microarray was used to compare differences in gene expression between tumor and normal vascular cells, and Expression Analysis Systematic Explorer was used to determine enrichment of gene ontology categories. Protein expression of select genes was confirmed using immunohistochemistry. Of the 1176 genes that were differentially expressed between tumor and normal vascular cells, 55 had a greater than fourfold increase in expression level. The extracellular matrix gene ontology category was increased while the ribosome gene ontology category was decreased. Fibroblast activation protein, secreted frizzledrelated protein 2, Janus kinase 3, and neutral sphingomyelinase 2 proteins localized to breast tumor endothelium as assessed by immunohistochemistry, showing significantly greater staining compared with normal tissue. These tumor endothelial marker proteins also exhibited increased expression in breast tumor vessels compared with that in normal tissues. Therefore, these genetic markers may serve as potential targets for the development of angiogenesis inhibitors.

Novel surface targets and serum biomarkers from the ovarian cancer vasculature

“…We have identified novel tumor-specific vasculature markers which appear promising for cancer serum diagnostics, molecular imaging and/or therapeutic targeting applications and warrant further clinical development. 4 Raphael sandaltzopoulos, 2 Andrew K. Godwin, 4 Nathalie scholler 1 and George Coukos 1, therapeutically to achieve destruction of established tumors or to prevent solid tumor growth by disrupting the tumor vasculature. Recent encouraging results with VEGF blockade have demonstrated the feasibility and efficacy of targeting tumor angiogenesis, [10][11][12][13] while vascular disrupting agents have produced dramatic results in preclinical models.…”

“…[1][2][3][4][5][6] Tumor vessels are enmeshed in a complex tissue, which includes tumor cells, stromal cells and leukocytes. Interactions with these cells and their products (growth factors, cytokines etc.)…”

Development of a Malignancy-Associated Proteomic Signature for Diffuse Large B-Cell Lymphoma

“…Although transcriptional profiling has matured as a diagnostic and prognostic research tool in pathology 11 and has been widely used to characterize malignancies as diverse as renal cell carcinoma, 12 prostate cancer, 13 metastatic bone cancer 14 and breast cancer, 15 proteinbased biomarkers may have potentially greater diagnostic power than genetic or transcriptional profiles because they may reflect disease-related alterations to tissue that are invisible to gene-based analyses. 16 Protein expression and activity are subject to additional layers of metabolic regulation over and above the regulation of gene expression.…”

Human transient receptor potential (TRP) channel expression profiling in carcinogenesis

“…In this regard, few papers have compared the expression profile of TEC versus their healthy counterpart defining several differentially expressed gene families (Bhati et al, 2008;Ghilardi et al, 2008). Nonetheless, although the expression and role of TRP channels in the vascular endothelium is well recognized and is subject of a number of recent reviews (Nilius and Droogmans, 2001;Yao and Garland, 2005), only recently TRPs have been recognized as major players in tumor vascularization (Fiorio Pla et al, 2012b;Fiorio Pla et al, 2012;Fiorio Pla and Gkika, 2013).…”