Abstract
Angiogenesis is a promising area of research that targets key therapeutic areas like cancer; wound healing, inflammatory diseases, etc. There is an increasing demand for screening of potential angiogenic and anti-angiogenic agents using sensitive, robust cell-based assays. We have developed a reporter vector containing cis-acting elements that respond to growth factors/angiogenic ligands for use in a cell-based luciferase reporter assay. We performed transient transfection of our reporter gene vector in MCF-7 cells to establish its application for screening of potential pro/antiangiogenic agents. Reporter gene transactivation studies with different concentrations of fetal bovine serum clearly indicated that the vector is functionally responsive to the angiogenic signals mediated by serum growth factors. We also used endostatin to inhibit transactivation and prove responsiveness to the anti-angiogenic agent. This vector is a promising tool for studying angiogenesis using cell-based reporter gene assays.
Keywords: Angiogenesis, endostatin, MCF-7, screening, serum, transfection, Cell Based, Reporter Gene Vector, inflammatory diseases, transactivation, extracellular matrix degradation, in vitro
Combinatorial Chemistry & High Throughput Screening
Title: Development of a Novel Reporter Gene Vector for Cell Based Angiogenic Studies
Volume: 14 Issue: 8
Author(s): Srinivasan Kokatam, Deepshikha Pandey, Ramkrishna Gupta, Rathu Krishna and Rajeev Soni
Affiliation:
Keywords: Angiogenesis, endostatin, MCF-7, screening, serum, transfection, Cell Based, Reporter Gene Vector, inflammatory diseases, transactivation, extracellular matrix degradation, in vitro
Abstract: Angiogenesis is a promising area of research that targets key therapeutic areas like cancer; wound healing, inflammatory diseases, etc. There is an increasing demand for screening of potential angiogenic and anti-angiogenic agents using sensitive, robust cell-based assays. We have developed a reporter vector containing cis-acting elements that respond to growth factors/angiogenic ligands for use in a cell-based luciferase reporter assay. We performed transient transfection of our reporter gene vector in MCF-7 cells to establish its application for screening of potential pro/antiangiogenic agents. Reporter gene transactivation studies with different concentrations of fetal bovine serum clearly indicated that the vector is functionally responsive to the angiogenic signals mediated by serum growth factors. We also used endostatin to inhibit transactivation and prove responsiveness to the anti-angiogenic agent. This vector is a promising tool for studying angiogenesis using cell-based reporter gene assays.
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Cite this article as:
Kokatam Srinivasan, Pandey Deepshikha, Gupta Ramkrishna, Krishna Rathu and Soni Rajeev, Development of a Novel Reporter Gene Vector for Cell Based Angiogenic Studies, Combinatorial Chemistry & High Throughput Screening 2011; 14 (8) . https://dx.doi.org/10.2174/138620711796504361
DOI https://dx.doi.org/10.2174/138620711796504361 |
Print ISSN 1386-2073 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5402 |
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