摘要
目的:探讨七氟醚调节血管内皮生长因子(VEGF)/血管内皮生长因子受体2(VEGFR2)信号通路对子宫内膜癌细胞增殖、凋亡和血管生成的影响。方法:将子宫内膜癌细胞HEC-1A分为对照组、七氟醚组、si-NC组、si-VEGF组、七氟醚+pc-NC组、七氟醚+pc-VEGF组;MTT、Edu法检测细胞增殖,Transwell实验检测细胞迁移和侵袭,流式细胞仪检测细胞凋亡率,Matrigel胶三维培养观察血管生成情况,qRT-PCR检测各组细胞中VEGF mRNA和VEGFR2 mRNA表达水平,Western blot检测细胞血管内皮生长因子A(VEGF-A)、血管内皮钙黏蛋白(VE-cadherin)、核蛋白(Ki-67)、半胱氨酸蛋白酶-3(Caspase-3)以及与VEGF/VEGFR2信号通路相关蛋白表达水平。结果:对照组细胞具有良好的管腔结构。与对照组相比,七氟醚组、si-VEGF组和七氟醚+pc-NC组HEC-1A细胞管腔结构明显被破坏,OD490(24h、48h)值和细胞增殖率、细胞迁移个数和侵袭个数、VEGF mRNA和VEGFR2 mRNA表达、VEGF-A、VE-cadherin、Ki-67、VEGF、VEGFR2蛋白表达显著降低,细胞凋亡率、Caspase-3蛋白表达显著升高(P<0.05),七氟醚组与si-VEGF组HEC-1A细胞各项检测指标均无差异(P>0.05);与七氟醚+pc-NC组相比,七氟醚+pc-VEGF组HEC-1A细胞管腔结构破坏程度显著减轻,OD490(24h、48h)值和细胞增殖率、迁移个数和侵袭个数、VEGF mRNA和VEGFR2 mRNA表达、VEGF-A、VE-cadherin、Ki-67、VEGF、VEGFR2蛋白表达显著升高,细胞凋亡率、Caspase-3表达显著降低(均P<0.05)。结论:七氟醚通过阻断VEGF/VEGFR2信号通路,抑制子宫内膜癌细胞增殖、血管生成,并促进其凋亡。
Objective:To investigate the impact the signaling pathway of vascular endothelial growth factor(VEGF)/vascular endothelial growth factor receptor 2(VEGFR2)regulated by sevoflurane on the proliferation,apoptosis,and angiogenesis of the endometrial cancer cells.Methods:The endometrial cancer cells HEC 1A were divided into control group,sevoflurane group,si NC group,si-VEGF group,sewoflurane+pc-NC group,and sevoflurane+pc-VEGF group.MTT and Edu methods were used t detect the proliferation of the endometrial cancer cells.T ranswell test was used to detect the migration and invasion of the cells.Flow cytometry was used to detect the apoptosis rate of the cells.Matrigel gel three dimensional culture was used to observe angiogenesis of the cells.The qRT-PCR was used to detect the expression levels of VEGF mRNA and VEGFR2 mRNA of the endometrial cancer cells in each group.w estern blot was used to detect the expression levels of vascular endothelial growth factor A(VEGF-A).vascular endothelial cad-herin(VE-cadherin),nuclear protein(Ki 67),cysteine protease-3(Caspase-3),and VEGF/VEGFR2 signaling path-way related proteins of the endometrial cancer cells.Results:The HEC 1A cells in the control group had good lumen structure.Compared with those of the HEC-1A cells in the control group,the lumen structure of the HEC-1A cells in the sevoflurane group,in the sirVEGF group,and in the sevoflurane+pc NC group had been disrupted significantly,the OD490 values in 24h and 48h,the cell proliferation rate,the numbers of cell migration and invasion,the expres-sion of VEGF mRNA and VEGFR2 mRNA,and the expression levels of VEGF-A.VE cadherin,Ki-67,VEGF,and VEGFR2 proteins of the HEC-1A cells in the sevoflurane group,in the si-VEGF group,and in the sevoflurane+pc-NC group had decreased significantly,and the apoptosis rate and the expression of Caspase-3 protein of the HEC-1A cells in the sevoflurane group.in the sir VEGF group,and in the sevoflurane+pe NC group had incrcased significantly(P<0.05).There were no stistically significant dffences in the detection indicators of the HEC-1A cells between the si-VEGF group and the sewoflurane group(P>0.05).Compared with those of the HEC 1A cells in the sevoflurane+pe-NC group,the damage degree of the lumen structure of the HEC-1A cells had decreases significantly,the OD490 walues in 24h and 48h,the proliferation rate,the numbers of migration and invasion,the expression levels of VEGF mRNA and VEGFR2 mRNA,and the expression levels of VEGF-A,VE cadherin,Ki-67,VEGF,and VEGFR2 pro-teins of the HEC-1A cells had increased significantly,the apoptosis rate and the expression of Caspase-3 protein had.decreased significantly(all P<0.05).Conclusion:Sevoflurane can inhibit the proliferation and angiogenesis of the en-dometrial cancer cells by blocking the VEGF/VEGFR2 signaling pathway,and can promote the apoptosis of the endo-metrial cancer cells.
作者
李正凯
冯玉
冯锋
王崇
薛金磊
LI Zhengkai;FENG Yu;FENG Feng;WANG Chong;XUE Jinlei(Zhengzhou Central Hospital Affiliated to Zhengzhou University,Henan Province,450007)
出处
《中国计划生育学杂志》
2023年第8期1777-1782,共6页
Chinese Journal of Family Planning
关键词
子宫内膜癌细胞
七氟醚
血管内皮生长因子
信号通路
血管生成
Endormetrial cancer cell
Sevoflurane
Vascular endothelial growth factor
Signaling pathway
Angiogene-sis