落新妇苷通过调节HIF-1α/VEGF轴抑制乳腺癌细胞的增殖、迁移和血管生成拟态形成

Astilbin inhibits proliferation,migration and vasculogenic mimicry formation of breast cancer cells by regulating HIF-1α/VEGF axis

目的:探究落新妇苷(AST)调节低氧诱导因子-1α(HIF-1α)/血管内皮生长因子(VEGF)轴对乳腺癌细胞增殖、迁移和血管生成拟态(VM)的影响。方法:用不同浓度的AST(0、5、25、50、100、150、200、300μmol/L)处理乳腺癌细胞MCF-7、MDA-MB-231,采用CKK-8法检测细胞活力。将MCF-7、MDA-MB-231细胞分为对照组、AST低剂量(AST-L)组、AST中剂量(AST-M)、AST高剂量(AST-H)组、AST-H+DMOG(HIF-1α/VEGF通路激活剂)组,EdU法检测AST处理对乳腺癌细胞增殖的影响,流式细胞术检测其对细胞凋亡的影响,Transwell小室实验检测其对细胞迁移、侵袭能力的影响,Matrigel管型形成实验检测其对细胞VM形成的影响,WB法检测对细胞中HIF-1α、VEGF、VE-cadherin、E-cadherin、N-cadherin、MMP-2表达的影响。结果:与0μmol/L AST相比,5、25、50、100、150、200、300μmol/L AST处理的细胞活力显著下降,呈剂量依赖性(P<0.05)。与对照组相比,AST-L、AST-M、AST-H组细胞EdU阳性率、细胞迁移数、细胞侵袭数、VM管腔数目、HIF-1α、VEGF、VE-cadherin、N-cadherin、MMP-2表达均显著下降,而细胞凋亡率、E-cadherin蛋白表达显著升高(均P<0.05);与AST-H组相比,AST-H+DMOG组细胞EdU阳性率、细胞迁移数、细胞侵袭数、VM管腔数目、HIF-1α、VEGF、VE-cadherin、N-cadherin、MMP-2表达均显著升高,而细胞凋亡率、E-cadherin蛋白表达均显著下降(均P<0.05)。结论:AST能抑制乳腺癌细胞增殖、迁移、侵袭和VM形成,促进凋亡,其作用机制可能与抑制HIF-1α/VEGF信号通路有关。

Objective:To explore the effects of astilbin(AST)on the proliferation,migration and vasculogenic mimicry(VM)formation in breast cancer cells by regulating the hypoxia-inducible factor-1α(HIF-1α)/vascular endothelial growth factor(VEGF)axis.Methods:Breast cancer cells(MCF-7 and MDA-MB-231)were treated with AST at different concentrations(0,5,25,50,100,150,200,300μmol/L),and the cell viability was detected using the CKK-8 assay.The MCF-7 and MDA-MB-231 cells were divided into control group,low-dose AST(AST-L),medium-dose AST(AST-M),high-dose AST(AST-H),and AST-H+DMOG(HIF-1α/VEGF pathway activator)groups.The effect of AST on cell proliferation was detected by EdU assay,the influence on apoptosis was detected using flow cytometry,the migration and invasion abilities of cells were examined using Transwell chamber assay,and the VM formation was analyzed using Matrigel tube formation assay.The expression of HIF-1α,VEGF,VE-cadherin,E-cadherin,N-cadherin and MMP-2 was detected by WB assay.Results:Compared with 0μmol/L AST,the viability of cells treated with 5,25,50,100,150,200,and 300μmol/L AST decreased obviously,which was in a dose-dependent manner(P<0.05).Compared with the control group,the AST-L,AST-M,and AST-H groups showed a significant decrease in EdU positive rate,cell migration number,cell invasion number,VM lumen number,and the protein expression of HIF-1α,VEGF,VE-cadherin,N-cadherin,and MMP-2,while the apoptosis rate and E-cadherin protein expression obviously increased(all P<0.05).Compared with the AST-H group,the AST-H+DMOG group demonstrated a significant increase in EdU positive rate,cell migration number,cell invasion number,VM lumen number,and the protein expression of HIF-1α,VEGF,VE-cadherin,N-cadherin,and MMP-2,while the apoptosis rate and E-cadherin protein expression significantly decreased(all P<0.05).Conclusion:AST can inhibit the proliferation,migration,invasion,and VM formation of breast cancer cells and promote apoptosis,which may be related to the inhibition of the HIF-1α/VEGF signaling pathway.