目的:研究柠檬苦素Toonaciliatin A对血管瘤内皮细胞(HemEC)增殖、凋亡的影响及潜在作用机制。方法:胶原蛋白酶A分离HemEC细胞,MTT法测定不同浓度Toonaciliatin A对HemEC细胞生存率的影响,流式细胞术测定HemEC细胞凋亡率,Western blot检测蛋白表达,qRT-PCR检测RNA的表达。结果:Toonaciliatin A能够抑制HemEC增殖,促进HemEC细胞凋亡;Toonaciliatin A抑制Tiam1蛋白表达,过表达Tiam1逆转了Toonaciliatin A对HemEC的抑制作用;Toonaciliatin A能够抑制VEGF和VEGFR2蛋白表达,过表达Tiam1逆转了Toonaciliatin A对VEGF和VEGFR2蛋白表达的抑制作用。结论:柠檬苦素类似物Toonaciliatin A通过靶向Tiam1基因抑制VEGF/VEGFR2通路抑制HemEC细胞增殖,促进细胞凋亡。
Objective:To investigate the effects of Limonin on proliferation and apoptosis of hemangioma endothelial cells(HemEC)and its underlying mechanism.Methods:HemEC cells were separated by collagenase A.Survival rate of HemEC treated with different dose of Toonaciliatin A was determined by MTT assay.Apoptotic rate of HemEC cells was measured by flow cytometry.Expression levels of proteins were detected by Western blot.Expression levels of RNA was detected by qRT-PCR.Results:Toonaciliatin A inhibited the proliferation and induced apoptosis in HemEC.Toonaciliatin A inhibited the expression levels of Tiam1 in HemEC.Overexpression of Tiam1 reversed the inhibitory effect of Toonaciliatin A on HemEC.Toonaciliatin A inhibited the expression of VEGF and VEGFR2.Overexpression of Tiam1 reversed the inhibitory effect of Toonaciliatin A on the expression of VEGF and VEGFR2.Conclusion:Toonaciliatin A-one of limonoids,inhibits the proliferation and reduced cell apoptosis in HemEC by targeting Tiam1 gene to inhibit the VEGF/VEGFR2 pathway.
Chinese Journal of Immunology
Hemangioma endothelial cells(HemEC)
T lymphoma invasion and metastasis inducing gene 1(Tiam1)
Vascular endothelial growth factor/vascular endothelial growth factor receptor 2(VEGF/VEGFR2)