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PTEN inhibits proliferation and VEGF expression in human umbilical vein endothelial ECV304 cells
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Abstract:
Objective: To investigate the effect of phosphatase and tensin hemology deleted on chromosome ten gene (PTEN) on proliferation, apoptosis, VEGF (vascular endothelial growth factor) and its receptor VEGFR1 expression in human umbilical vein endothelial cell (HUVEC) line ECV304. Methods: Recombinant adenovirus containing green fluorescent protein (GFP) and PTEN (AdPTENGFP) or empty vector (AdGFP) were transfected into ECV304 cells; proliferation and apoptosis of ECV304 cells were measured by MTT assay, Hoechst3342 staining and flow cytometry, respectively; PTEN, VEGF, VEGFR1 mRNA expression levels in AdPTENGFPtransfected ECV304 cells were examined by quantitative PCR; and VEGF protein level in ECV304 cell supernatant was detected by ELISA. Chick chorioallantoic membrane (CAM) assay was used to study the effect of PTEN on angiogenesis. Results: AdPTENGFP transfection significantly inhibited the proliferation and induced apoptosis of ECV304 cells and the inhibitory rate and apoptotic rate were (50.38±5.42)% and (73.3±5.3)% at 5 d. VEGF and VEGFR1 mRNA expression levels were (13.40±1.32)% and (46.12±5.20)% of untransfected group after transfected with AdPTENGFP at MOI=100 in ECV304 cells. Furthermore, CAM assay results showed that AdPTENGFP transfection inhibited CAM angiogenesis in vivo. Conclusion: PTEN can inhibit the growth of and promote apoptosis of human umbilical vein endothelial ECV304 cells, which might be related to the downregulation of VEGF/VEGFR1 expression and the resulting angiogenesis inhibition.
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Project support by the Key Research Program of Science and Technolgy in Baoding (No. 11ZF003)
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