摘要
目的 探讨藁本内酯对胃癌细胞增殖、凋亡和血管生成的影响及其作用机制.方法 培养人胃癌细胞NCI-N87,细胞计数试剂盒-8(CCK-8)法检测不同浓度Ligustilide对NCI-N87细胞存活率的影响;根据实验要求将细胞分为对照组,藁本内酯低、中、高浓度组(藁本内酯-L、M、H组),藁本内酯-H+DMOG(HIF-1α激活剂)组;采用EDU实验检测各组NCI-N87细胞增殖能力;流式细胞术检测各组NCI-N87细胞凋亡;酶联免疫吸附法(ELISA)检测各组NCI-N87细胞分泌血管内皮生长因子A(VEGFA)水平;体外管形成实验检测各组细胞血管生成情况;蛋白免疫印迹(Western blot)检测各组NCI-N87细胞缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)、细胞增殖核抗原(Ki67)、裂解的半胱天冬酶-3(cleaved-Caspase3)、半胱天冬酶-3(Caspase3)蛋白表达.结果 与对照组相比,藁本内酯-L、M、H组NCI-N87细胞增殖能力、培养液中VEGFA水平、血管生成能力、细胞HIF-1α、VEGF、Ki67蛋白表达显著降低,细胞凋亡率以及cleaved-Caspase3/Caspase3蛋白表达显著升高,且存在浓度依赖性(P<0.05);与藁本内酯-H组相比,藁本内酯-H+DMOG组可部分逆转藁本内酯-H组对NCI-N87细胞行为及以上蛋白表达的影响(P<0.05).结论 藁本内酯可通过调节HIF-1α/VEGF信号通路抑制胃癌细胞增殖、血管生成,并诱导其凋亡.
Abstract
Objective To investigate the effect and mechanism of Ligustilide on the proliferation,apoptosis and angiogenesis of gastric cancer cells.Methods Human gastric cancer cells NCI-N87 were cultured,and the effect of Ligustilide on the survival of NCI-N87 cells was detected by cell counting Kit 8(CCK-8).The cells were divided into the control group,low,medium and high doses of Ligustilide groups(Ligustilide-L,M and H groups),and Ligustilide-H+DMOG(HIF-1α activator)group.The proliferation of NCI-N87 cells was detected by EDU assay.The apoptosis of NCI-N87 cells was detected by flow cytometry.The level of vascular endothelial growth factor A(VEGFA)secreted by NCI-N87 cells was detected by enzyme-linked immunosorbent assay(ELISA).Angiogenesis of cells was detected by tube formation in vitro.Western blot was applied to detect the expression of hypoxia-induced factor-1α(HIF-1α),vascular endothelial growth factor(VEGF),cell proliferation-associated nuclear antigen(Ki67),cleaved caspase-3,and caspase-3 in NCI-N87 cells.Results Compared with the control group,the proliferation,VEGFA level,angiogenesis,and the expression of HIF-1α,VEGF and Ki67 in NCI-N87 cells were significantly decreased in the Ligustilide-L,M and H groups,but the cell apoptosis rate and cleaved-Caspase3/Caspase3 expression were significantly increased in a concentration-dependent manner(P<0.05).Compared with Ligustilide-H group,Ligustilide-H+DMOG group partially reversed the effect of Ligustilide-H group on the behavior and above protein expression of NCI-N87 cells(P<0.05).Conclusion Ligustilide can inhibit the proliferation and angiogenesis,and induce apoptosis of gastric cancer cells by regulating the HIF-1α/VEGF pathway.
基金项目
广东省中医药局科研项目(202305042253436670)
维普
万方数据