西罗莫司对TIE2-L914F突变导致的静脉畸形血管内皮细胞增殖和凋亡的影响及其体外机制探究  

Effects of sirolimus on cell proliferation and apoptosis of vascular endothelial cells of venous malformation induced by TIE2-L914F mutation and its mechanism in vitro

作　　者：施磊 孙圆圆 甘露 王新婧 王海宝[1] Shi Lei;Sun Yuanyuan;Gan Lu;Wang Xinjing;Wang Haibao(Dept of Radiology,The First Affiliated Hospital of Anhui Medical University,Hefei 230022;Dept of Radiology,Huainan Oriental Hospital Group General Hospital,Huainan 232038;Dept of Radiology,Anhui Maternal and Child Health Hospital,Hefei 230001)

机构地区：[1]安徽医科大学第一附属医院放射科,合肥230022 [2]淮南东方医院集团总医院放射科,淮南232038 [3]安徽省妇幼保健院放射科,合肥230001

出　　处：《安徽医科大学学报》2023年第4期561-567,共7页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金(编号:1808085MH228)。

摘　　要：目的 探究西罗莫司(SIR)对TIE2-L914F突变导致的静脉畸形(VM)血管内皮细胞增殖和凋亡的影响及其潜在分子机制。方法 干预人脐静脉内皮细胞(HUVEC)中TEK受体酪氨酸激酶(TIE2)的表达以构建VM血管内皮细胞模型(TIE2-L914F组)。之后将部分VM血管内皮细胞模型暴露于1 000 ng/ml SIR 48 h(TIE2-L914F+SIR组),采用MTT和流式细胞术检测VM血管内皮细胞增殖和凋亡。采用qRT-PCR和Western blot检测CXC趋化因子配体(CXCL)1和CXCR2的mRNA及蛋白表达。结果 与TIE2-L914F组细胞相比,TIE2-L914F+SIR组细胞增殖活力受到抑制,凋亡率升高,差异有统计学意义(P<0.05)。CXCL1在TIE2-L914F细胞中表达升高,在SIR处理后降低,差异有统计学意义(P<0.05)。与TIE2-L914F+pcDNA3.1组比较,TIE2-L914F+pcDNA-CXCL1组细胞增殖活力提高,凋亡率降低;而与TIE2-L914F+pcDNA-CXCL1组比较,TIE2-L914F+pcDNA-CXCL1+SIR组细胞增殖活力被抑制,凋亡率提高,差异有统计学意义(P<0.05)。此外,与TIE2-L914F组比较,TIE2-L914F+SIR组CXCR2表达降低(P<0.05)。结论 SIR能够抑制TIE2-L914F突变导致的VM血管内皮细胞增殖,诱导细胞凋亡,抑制CXCL1/CXCR2表达。Objective The purpose of this study was to investigate the effects of sirolimus(SIR)on proliferation and apoptosis of vascular endothelial cells of venous malformation(VM)caused by mutations in TIE2-L914F and its potential molecular mechanism.Methods The expression of TEK receptor tyrosine kinase(TIE2)in human umbilical vein endothelial cell(HUVEC)was interfered to construct vascular endothelial cells of VM model(TIE2-L914F group).Subsequently part of vascular endothelial cells of VM was exposed to 1000 ng/ml SIR for 48 h(TIE2-L914F+SIR group),and the proliferation and apoptosis of vascular endothelial cells of VM were detected by MTT and flow cytometry.The mRNA and protein expressions of CXCL1 and CXCR2 were detected by qRT-PCR and Western blot.Results Compared with the cells in TIE2-L914F group,the proliferation activity of the cells in TIE2-L914F+SIR group was inhibited,and the apoptosis rate increased(P<0.05).The expression of CXCL1 increased in TIE2-L914F cells but decreased after SIR treatment(P<0.05).Compared with TIE2-L914F+pcDNA3.1 group,the cell proliferation activity increased but apoptosis rate decreased in TIE2-L914F+pcDNA-CXCL1 group.Compared with TIE2-L914F+pcDNA-CXCL1 group,cell proliferation activity was inhibited but apoptosis rate increased in TIE2-L914F+pcDNA-CXCL1+SIR Group(P<0.05).In addition,compared with TIE2-L914F group,CXCR2 expression decreased in TIE2-L914F+SIR group(P<0.05).Conclusion SIR inhibits VM cell proliferation,induces cell apoptosis of vascular endothelial cells of VM,and inhibits the expression of CXCL1/CXCR2.

关 键 词：西罗莫司 CXCL1/CXCR2 静脉畸形 增殖 凋亡 

分 类 号：R654.3[医药卫生—外科学]