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U87 glioma cell supernatant induces angiogenesis of CD133+ endothelial cells and possible mechanism
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Abstract:
Objective : To investigate the angiogenesis of CD133+ endothelial cells induced by glioma U87 cell supernatant and the possible mechanisms. Methods: CD133+ cells were sorted from the cord blood by magnetic activated cell sorting system, and were further induced to CD133+ endothelial cells. CD133+ endothelial cells were indetified by confocal microscopy and treated with U87 cell supernatant (DMEM used as control), and their angiogenesis and migration were examined by in vitro angiogenesis assay and Transwell assay, respectively. VEGFR-1, VEGFR-2 and MMP-9 (matrix metalloproteinase 9) expressions were determined by Western blotting analysis, and MMP-9 activity was examined by gelatin zymography assay. Results: After in vitro cultured for 14 d, the cord blood CD133+ cells showed positive expression of Dil-ac-LDL and FITC-UEA-1 and had the features of endothelial cells, so they were named CD133+ endothelial cells. The supernatant of U87 glioma cells induced tube formation of CD133+ endothelial cells and increased migration of CD133+ endothelial cells. U87 cell supernatant induced angiogenesis (\[40.7±3.3\] vs \[21±2.3\], P<0.05), increased migration (\[0.60±0.04\] vs \[0.27±002\], P<0.05), up-regulated VEGFR-2 and MMP-9 expressions (P<0.05) in CD133+ endothelial cells, while had minimal effect on VEGFR-1 expression. Moreover, U87 cell supernatant enhanced MMP-9 activity of CD133+ endothelial cells. Conclusion: Glioma U87 cell supernatant can induce angiogenesis of CD133+ endothelial cell via up-regulating VEGFR-2 and MMP-9 expressions.
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Project Supported:
Project supported by the Medical Major Talent Program of Jiangsu Province（No.RC2007061）
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