Combretastatin A-4氨基糖类衍生物CPU-XT-008抑制血管内皮细胞增殖的分子机制
Molecular mechanisms of combretastatin A-4 amino sugar derivative CPU-XT-008 on inhibiting proliferation of vascular endothelial cells
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摘要: 以人脐静脉内皮细胞(HUVEC)为模拟肿瘤血管内皮细胞的实验模型，研究combretastatin A-4(CA-4)的氨基糖类衍生物CPU-XT-008对HUVEC的增殖、周期分布、微管蛋白聚合以及关键周期调控蛋白表达的影响。采用MTT法检测CPU-XT-008对HUVEC增殖的影响；采用流式细胞仪检测CPU-XT-008对HUVEC周期分布的影响；采用免疫荧光染色检测CPU-XT-008对HUVEC凋亡及β-tubulin微管蛋白的聚合变化；采用Western blot检测CPU-XT-008对周期蛋白Cyclin B1，Cdc2和微管蛋白β-tubulin的影响。结果表明，CPU-XT-008能以微管蛋白为靶点抑制β-tubulin微管蛋白聚合，通过上调HUVEC的细胞周期蛋白Cyclin B1水平表达，下调细胞周期依赖性激酶Cdc2蛋白水平的表达从而引发细胞G2/M期阻滞，抑制细胞增殖并诱导细胞凋亡。
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关键词:
- combretastatin A-4 /
- CPU-XT-008 /
- 氨基糖类衍生物 /
- 人脐静脉内皮细胞 /
- 微管蛋白 /
- 细胞周期
Abstract: Taking human umbilical vein endothelial cells(HUVEC)as experimental model which can simulate tumor-derived vascular endothelial cells, the effects of CPU-XT-008, an amino sugar derivative of combretastatin A-4(CA-4), on HUVEC proliferation, cell cycle distribution, tubulin polymerization and the key regulatory protein of cell cycle were studied. The effect of CPU-XT-008 on the proliferation of HUVEC was determined by MTT assay. The cell cycle distribution caused by CPU-XT-008 was detected by flow cytometry. Immunofluorescence was used to detect apoptosis and tubulin polymerization. The expressions of Cyclin B1, Cdc2 and β-tubulin were detected by Western blotting. Results demonstrated that CPU-XT-008 could target tubulin and inhibit the polymerization of β-tubulin, and it could lead to G2/M cell cycle arrest in HUVEC by up-regulating Cyclin B1 expression and down-regulating Cdc2 and p-Cdc2 expression, which resulted in inhibiting the proliferation of HUVEC and inducing its apoptosis. -
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