Subtractive transcriptomics: establishing polarity drives in vitro human endothelial morphogenesis

Cancer Research
David A GlesneDiane J Rodi

Abstract

Although investigations of mature normal and tumor-derived capillaries have resulted in characterization of these structures at the phenotypic level, less is known regarding the initial molecular cues for cellular assembly of endothelial cells into human capillaries. Here, we employ a novel combination of microenvironmental manipulation and microarray data filtration over narrowly delineated temporal data series to identify the morphogenesis component apart from the proliferation component, as pooled human microvascular-derived endothelial cells are induced to form capillary-like structures in vitro in a murine tumor-derived matrix. The 217 morphogenesis-specific genes identified using this subtractive transcriptomics approach are mostly independent of the angiogenic proteins currently used as therapeutic targets for aberrant angiogenesis. Quantitative real-time PCR was used to validate 20% of these transcripts. Immunofluorescent analysis of proliferating and tube-forming cells validates at the protein level the morphogenesis-specific expression pattern of 16 of the 217 gene products identified. The transcripts that are selectively up-regulated in tube-forming endothelial cells reveal a temporal expression pattern of genes prim...Continue Reading

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Citations

May 29, 2008·Clinical and Experimental Pharmacology & Physiology·Lydia FinneyDavid Glesne
Feb 7, 2007·Proceedings of the National Academy of Sciences of the United States of America·Lydia FinneyDavid Glesne
Oct 2, 2014·Canadian Journal of Physiology and Pharmacology·Hongyan ZhouShao Bo Su
Jun 25, 2009·Scandinavian Journal of Clinical and Laboratory Investigation·Johanna M MattssonHannu Koistinen
Nov 18, 2011·Integrative Biology : Quantitative Biosciences From Nano to Macro·Magdalena A CichonDerek C Radisky
Feb 26, 2016·Current Opinion in Hematology·Toni M AntalisMarguerite S Buzza
Jan 13, 2009·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Sofie MellbergLena Claesson-Welsh
Aug 23, 2012·Physiological Genomics·Liang-Hui ChuJoel S Bader

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