HIF-1α靶向SP1促进食管鳞状细胞癌转移及血管生成的机制研究

Mechanism of HIF-1αtargeting SP1 to promote metastasis and angiogenesis of esophageal squamous cell carcinoma

目的:探究HIF-1α对食管鳞状细胞癌转移及血管生成的作用及其可能的机制。方法:RT-PCR检测细胞HIF-1α和SP1 mRNA表达;Western blot检测细胞HIF-1α和SP1蛋白表达;CCK-8检测细胞活力;Transwell实验检测细胞迁移和侵袭;体外血管生成实验检测人脐静脉血管内皮细胞HUVEC血管形成能力;ChIP-PCR实验检测HIF-1α和SP1启动子的结合。结果:与人食管上皮细胞HEEC相比,在食管鳞状细胞癌细胞Ec109、KYSE30、KYSE150和KYSE410中HIF-1α和SP1 mRNA和蛋白表达显著上调(P<0.05),其中Ec109细胞变化最为明显。与sh-NC组相比,sh-HIF-1α组Ec109细胞活力、迁移和侵袭能力、HUVEC细胞管道数目显著下调(P<0.05);与oe-NC组相比,oe-HIF-1α组Ec109细胞活力、迁移和侵袭能力、HUVEC细胞管道数目显著上调(P<0.05)。HIF-1α靶向结合SP1启动子区。与sh-NC组相比,sh-HIF-1α组Ec109细胞SP1 mRNA和蛋白表达、细胞活力、迁移和侵袭能力、HUVEC细胞管道数目显著下调(P<0.05);与sh-HIF-1α+oe-NC组相比,sh-HIF-1α+oe-SP1组Ec109细胞SP1 mRNA和蛋白表达、细胞活力、迁移和侵袭能力、HUVEC细胞管道数目显著上调(P<0.05)。结论:HIF-1α通过靶向SP1促进食管鳞状细胞癌转移及血管生成。

Objective:To explore the effect of HIF-1αon metastasis and angiogenesis of esophageal squamous cell carcinoma and its possible mechanism.Methods:RT-PCR was used to detect HIF-1αand SP1 mRNA expression in cells.Western blot was used to detect HIF-1αand SP1 protein expression.CCK-8 was used to detect cell viability.Transwell test was used to detect cell migration and invasion.Angiogenesis test in vitro was used to detect the angiogenesis ability of human umbilical vein blood vessels endothelial cells HUVEC.ChIP-PCR test was used to detect the binding of HIF-1αand SP1 promoter.Results:Compared with human esophageal epithelial cells HEEC,the expressions of HIF-1αand SP1 mRNA and protein were significantly up-regulated in esophageal squamous cell carcinoma cells Ec109,KYSE30,KYSE150,and KYSE410(P<0.05),among which Ec109 cells had the most obvious changes.Compared with the sh-NC group,the viability,migration and invasion ability of Ec109 cells in the sh-HIF-1αgroup and the number of HUVEC cell channels were significantly downregulated(P<0.05).Compared with the oe-NC group,the cell viability,migration and invasion ability,and number of HUVEC cell channels of Ec109 cells in the oe-HIF-1αgroup were significantly increased(P<0.05).HIF-1αtargets the SP1 promoter region.Compared with the sh-NC group,the expression of SP1 mRNA and protein,cell viability,migration and invasion ability,and HUVEC cell channels of Ec109 cells in the sh-HIF-1αgroup were significantly down-regulated(P<0.05).Compared with sh-HIF-1α+oe group,the expression of SP1 mRNA and protein,cell viability,migration and invasion ability,and the number of HUVEC cell channels of Ec109 cells in the sh-HIF-1α+oe-SP1 group were significantly increased(P<0.05).Conclusion:HIF-1αpromotes esophageal squamous cell carcinoma metastasis and angiogenesis by targeting SP1.