【摘要】 目的探讨芹菜素对人乳腺癌细胞血管内皮生长因子(VEGF)表达的影响及作用机制。方法采用四甲基偶氮唑盐(MTT)法检测芹菜素对人乳腺癌细胞系MDA-MB-231细胞增殖的影响;应用酶联免疫吸附试验(ELISA)检测VEGF的分泌水平;应用逆转录-聚合酶链反应(RT-PCR)检测VEGF mRNA表达水平;采用双荧光素酶系统检测VEGF转录活性;通过转入pCEP4-HIF-1α观察缺氧诱导因子-1α(HIF-1α)逆转芹菜素抑制VEGF转录活性的作用;应用Western blot检测芹菜素对HIF-1α、p-AKT、p-ERK1/2和p53表达的时间依赖性和剂量依赖性效应。结果芹菜素对MDA- MB-231细胞无明显增殖抑制效应;芹菜素降低了VEGF分泌水平、mRNA表达水平以及VEGF的转录活性,并且芹菜素对VEGF转录活性的抑制作用能够通过转入HIF-1α而逆转。芹菜素抑制HIF- 1α和p-AKT的表达,诱导p53的表达,而对p-ERK1/2的表达没有影响。结论芹菜素能够抑制乳腺癌细胞VEGF的表达,这种效应可能通过降低HIF-1α的表达来实现。更多还原

【Abstract】 Objective To study the effects and mechanism of apigenin on the expression of vascular endothelial growth factor (VEGF) in human breast cancer MDA-MB-231 cells. Methods MDA-MB-231 cells were used as the study object. MTT assay was used to detect the effect of apigenin on the cell viability. ELISA was used to determine the protein level of VEGF. RT-PCR was used to detect VEGF at mRNA level. A double luciferase system was used to measure the transcription activity of VEGF. pCEP4-HIF-1αwas transferred to explore the reversing effect of HTF-1αon the inhibitory effect of apigenin on the transcription activity of VEGF. Western blotting was used to detect the time-dependent and dose-dependent effect of apigenin on HIF-1α,p-AKT,p-ERK,and p53 expression at protein level. Results Apigenin had no visible inhibitory effect on cell viability. Apigenin reduced the secretion of VEGF, mRNA levels of VEGF and transcription activity of VEGF. Furthermore, the inhibitory effect of apigenin on the transcription activity of VEGF could be reversed by transferring pCEP4-HIF-1αinto the cells. Additionally, apigenin inhibited the expression of HIF-1αand p-AKT, induced the expression of p53, but had no effect on the expression of p-ERK1/2. Conclusion Apigenin can inhibit VEGF expression in breast cancer cells, and this effect may be achieved through decreasing the expression of HIF-1α.更多还原