USP11调控VEGF表达及血管形成促进肝癌细胞的侵袭和转移

USP11 promotes invasion and metastasis of hepatocellular carcinoma cells through regulating VEGF expression and angiogenesis

目的:探究去泛素化酶11(USP11)调控VEGF和血管形成促进肝癌细胞侵袭和转移能力的机制。方法:运用Real-time PCR实验检测USP11对肝癌细胞中VEGF基因表达水平的影响;应用Elisa实验检测USP11对肝癌细胞分泌VEGF蛋白的影响;利用对照组和USP11敲低组肝癌细胞分泌上清分别处理血管内皮细胞HUVECs,运用Transwell基质胶实验检测血管内皮细胞的迁移能力;运用CCK-8实验检测血管内皮细胞的增殖能力;运用小管形成实验检测血管内皮细胞的成管能力。结果:敲低USP11可降低肝癌细胞中VEGF的基因表达水平(P<0.01),并抑制肝癌细胞分泌VEGF蛋白(P<0.001);与对照组相比,敲低USP11的肝癌细胞分泌上清处理血管内皮细胞,血管内皮细胞的增殖能力减弱(P<0.01),成管能力也远低于对照组;两组肝癌细胞和血管内皮细胞共培养,基质胶实验表明,敲低USP11组,血管内皮细胞的迁移能力降低(P<0.01)。结论:USP11可以增加VEGF的基因转录和蛋白表达水平,增强血管内皮细胞的增殖、迁移和成管能力,进而促进肝癌细胞的侵袭和转移。

Objective:To explore the mechanism of deubiquitination enzyme 11(USP11)regulating VEGF and angiogenesis to promote invasion and metastasis of hepatocellular carcinoma(HCC)cells.Methods:Real-time PCR was used to detect the effect of USP11 on the expression level of VEGF gene in HCC cells.The effect of USP11 on the secretion of VEGF protein in HCC cells was detected by Elisa.Vascular endothelial cells HUVECs were treated with supernatant secreted by HCC cells in control and USP11 knockdown group respectively,and transwell matrix gel assay was used to detect the migration ability of vascular endothelial cells.The proliferation ability of vascular endothelial cells was detected by CCK-8 assay and the angiogenesis ability of vascular endothelial cells was detected by tubule formation assay.Results:Knockdown of USP11 could decrease the gene expression level of VEGF in HCC cells(P<0.01),and inhibit the secretion of VEGF protein(P<0.001).Compared with the control group,the proliferation ability of vascular endothelial cells was weaker in USP11 knocked down group(P<0.01),and the angiogenesis ability was much lower than that of the control group.HCC cells and vascular endothelial cells were co-cultured in the two groups.Matrix gel experiment showed that the migration ability of vascular endothelial cells was decreased in USP11 knocked down group(P<0.01).Conclusion:USP11 increases the gene transcription and protein expression level of VEGF,enhances the proliferation,migration and angiogenesis ability of vascular endothelial cells,thus promoting the invasion and metastasis of HCC cells.