【摘要】 目的研究血管内皮生长因子(VEGF)对上皮性卵巢癌细胞株AO体外侵袭能力和对细胞中基质金属蛋白酶-2(MMP-2),金属蛋白酶组织抑制剂-2(TIMP-2)表达的影响,探讨VEGF在卵巢癌浸润转移中的作用及可能机制。方法以处理卵巢癌细胞株AO,采用Boyden小室侵袭实验检测AO细胞体外侵袭能力,采用RT-PCR和Western免疫印迹检测AO细胞MMP-2的mRNA及蛋白表达情况,采用DQ明胶荧光检测法分析MMP-2酶活性。结果VEGF能双相调节卵巢癌细胞AO的体外侵袭能力及细胞中MMP-2的分泌及活性,低浓度(20￣40ng/mL)时明显增强癌细胞体外侵袭能力,上调MMP-2的分泌及活性,并随着VEGF的浓度增加该作用增强;但高浓度时(80ng/mL),其体外侵袭能力减弱,MMP-2的活性下降。MMP抑制剂BB3103能降低VEGF刺激后卵巢癌细胞AO的体外侵袭能力。此外,VEGF不影响卵巢癌细胞AO中MMP-2,TIMP-2的基因表达。结论VEGF促进卵巢癌侵袭转移,其作用机制的关键环节是促进MMP-2的分泌与活化。更多还原

【Abstract】 [Objective] To investigate the effect of vascular endothelial growth factor (VEGF) on the invasive ability and the expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitors of matrix metalloproteinases (TIMP-2) in ovarian cancer cell lines AO and associated mechanism. [Methods] AO Cells were cultured for 48 hours and then treated with VEGF at different concentrations for additional 18 hours. The invasive ability of AO cells was analyzed with Boyden chamber invasion assay. Reverse transcriptase ploymerase chain reaction and western blot were used to detect the expression of MMP-2 secretion in conditional medium. The activity of MMP-2 was analyzed by fluorogenic DQ gelatin. [Results] VEGF, at 20~40 ng/mL, enhanced the invasive ability and induced secretion and activation of pro-MMP-2 in conditioned medium in a concentration dependent manner. However, gelatinolytic activity reached the maximum upon VEGF treatment at 40 ng/ml and decreased at higher-concentration VEGF treatment (80 ng/mL), However. The expression of MMP-2 and TIMP-2mRNA in AO had no response to VEGF stimulation. [Conclusion] VEGF may regulate ovarian cancer invasion and migration and induction of MMP-2 by VEGF is a key component of VEGF-induced ovarian cancer cell invasion.更多还原