基于VEGF/VEGFR信号通路研究双氢青蒿素对食管癌细胞生物学行为的影响及其机制

Exploration on the effects and mechanism of dihydroartemisinin on biological behaviors of esophageal cancer cells based on VEGF/VEGFR signaling pathways

目的:基于血管内皮生长因子(VEGF)/VEGF受体(VEGFR)信号通路探讨双氢青蒿素(DHA)对食管癌细胞生物学行为的影响。方法:将人食管癌Eca-109细胞分为对照组、DHA组、DHA+pcNC组和DHA+pc VEGF组,使用CCK8法检测细胞增殖情况,qRT-PCR检测细胞增殖基因、VEGF和VEGFR2 mRNA水平,流式细胞仪检测细胞凋亡率,Transwell实验法检测细胞侵袭情况,Western blot检测细胞凋亡、侵袭相关蛋白表达。结果:与对照组相比,DHA组Eca-109细胞中的VEGF、VEGFR2 mRNA水平降低,细胞增殖活性抑制率、细胞凋亡率、单个视野细胞侵袭数目和cleaved-Caspase-3、Bax、MMP-2、MMP-9蛋白表达升高,Ki-67、PCNA、Bcl-2蛋白表达降低(P<0.05)。与DHA+pcNC组相比,DHA+pc VEGF组Eca-109细胞中的VEGF、VEGFR2 mRNA水平升高,细胞增殖活性抑制率、细胞凋亡率、单个视野细胞侵袭数目和cleaved-Caspase-3、Bax、MMP-2、MMP-9蛋白表达降低,Ki-67、PCNA、Bcl-2蛋白表达升高(P<0.05)。结论:DHA对食管癌Eca-109细胞增殖、侵袭有抑制作用,对细胞凋亡有促进作用,可能与抑制VEGF/VEGFR信号通路活化有关。

Objective:To explore the effects of dihydroartemisinin(DHA)on biological behaviors of esophageal cancer cells based on vascular endothelial growth factor(VEGF)/VEGF receptor(VEGFR)signaling pathways.Methods:Human esophageal cancer Eca-109 cells were divided into control group,DHA group,DHA+pcNC group and DHA+pc VEGF group.The cells proliferation was detected by CCK8,levels of cell proliferation gene,VEGF and VEGFR2 mRNA were detected by qRT-PCR,cells apoptosis rate was detected by flow cytometry,cell invasion was detected by Transwell assay,and expressions of apoptosis and invasion-related proteins were detected by Western blot.Results:Compared with control group,mRNA levels of VEGF and VEGFR2 in Eca-109 cells were decreased in DHA group,inhibition rate of cells proliferation activity,apoptosis rate,number of invasion cells under single visual field and expressions of cleaved-Caspase-3,Bax,MMP-2 and MMP-9 proteins were increased,and expressions of Ki-67,PCNA,Bcl-2 proteins were decreased(P<0.05).Compared with DHA+pcNC group,mRNA levels of VEGF and VEGFR2 in Eca-109 cells were increased in DHA+pc VEGF group,inhibition rate of cells proliferation activity,apoptosis rate,number of invasion cells under single visual field and expressions of cleaved-Caspase-3,Bax,MMP-2 and MMP-9 proteins were decreased,and expressions of Ki-67,PCNA,Bcl-2 proteins were increased(P<0.05).Conclusion:DHA can inhibit proliferation and invasion of Eca-109 cells,and promote cells apoptosis,which may be related to inhibiting the activation of VEGF/VEGFR signaling pathways.