平阳霉素对静脉畸形细胞增殖及凋亡的作用研究

Effect of pingyangmycin on the proliferation and apoptosis of venous malformation cells

目的:探讨平阳霉素对静脉畸形细胞增殖及凋亡的影响。方法:在人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)中过表达TIE2-L914F与TIE2-WT基因从而构建静脉畸形细胞模型和野生型细胞模型组,HUVEC为Control组。采用CCK-8检测平阳霉素对静脉畸形细胞增殖活性的影响。Hoechst 33342染色及流式细胞计数法检测平阳霉素对静脉畸形细胞凋亡的影响。Western Blot检测凋亡相关蛋白Caspase-3、Caspase-9、Bax、Bcl-2及P53的表达水平。结果:CCK-8结果显示平阳霉素可以明显抑制静脉畸形细胞的增殖活性,且呈时间依赖性与剂量依赖性(P<0.05)。Hoechst 33342染色及流式细胞计数结果显示平阳霉素可明显促进静脉畸形细胞凋亡(P<0.05)。Western Blot结果显示经平阳霉素处理后,促凋亡蛋白Caspase-3与Caspase-9的活化水平上调,Bax及P53表达上调,抑凋亡蛋白Bcl-2表达下调(P<0.05)。结论:平阳霉素可明显抑制静脉畸形细胞的增殖活性,可能通过线粒体途径促进凋亡,从而发挥治疗作用。

Objective:To investigate the effect of pingyangmycin(PYM)on the proliferation and apoptosis of venous malformation cells.Methods:TIE2-L914 F and TIE2-WT genes were overexpressed in human umbilical vein endothelial cells(HUVEC)to construct a venous malformation cell model and a wild type cell model.HUVEC was a control group.CCK-8 was used to detect the effect of PYM on the proliferation of venous malformation cells.Hoechst 33342 staining and flow cytometry were used to detect the effect of PYM on the apoptosis of venous malformation cells.The apoptotic protein expression levels of caspase-3,caspase-9,Bax,Bcl-2 and P53 were detected by Western Blot.Results:CCK-8 results showed that PYM could significantly inhibit the proliferation of venous malformation cells in a time-and dose-dependent manner(P<0.05).Hoechst 33342 staining and flow cytometry showed that PYM could significantly promote the apoptosis of venous malformation cells(P<0.05).Western Blot results showed that the activation level of Caspase-3 and Caspase-9 pro-apoptosis-related proteins were up-regulated,the expression level of Bax and P53 were up-regulated,while the expression of Bcl-2 was down-regulated(P<0.05).Conclusion:PYM can significantly inhibit the proliferation of venous malformation cells and may promote apoptosis through mitochondrial pathway,thus playing a therapeutic role.