Mechanism of miR-204 inhibiting angiogenesis of gastric cancer by regulating ZEB1
Objective To investigate the underlying mechanism of miR-204 inhibiting angiogenesis of gastric cancer by regulating zinc finger E-box-binding protein 1(ZEB1).Methods The expression levels of miR-204 and ZEB1 in tumors and adjacent tissues of clinical gastric cancer patients were analyzed by using TCGA database and immunohistochemical staining.The expression levels of miR-204 and ZEB1 in gastric cancer patients with different clinicopathological stages and lymphatic metastasis,and their correlation with overall survival(OS)were analyzed by using the UALCAN database.Angiogenesis assay was used to detect the changes of angiogenesis ability of human umbilical vein endothelial cells(HUVEC)after cultured in conditioned medium with miR-204 overexpression and knockdown in HGC-27 cells.The effects of miR-204 on the protein expression levels of ZEB1,VEGFR1 and VEGFR2 in HGC-27 cells were detected by western blot.Results TCGA database and immunohistochemical staining results showed that compared with adjacent tissues,the expression levels of miR-204 in gastric cancer tissues was significantly decreased(P＜0.001),while the expression levels of ZEB1 was significantly upregulated(P＜0.001),and Pearson correlation analysis result showed a negative correlation between miR-204 and ZEB1 expression levels(r=-0.827,P＜0.001).The results of UALCAN database analysis showed that the expression levels of miR-204 in gastric cancer patients with different clinicopathological stages and lymphatic metastasis were significantly decreased than that in normal tissues(F=17.817,18.538;all P＜0.001),and was positively correlated with OS in gastric cancer patients(r=0.547,P＜0.05),while the expression levels of ZEB1 in gastric cancer patients with different clinicopathological stages and lymphatic metastasis were significantly increased and higher than that in normal tissues(F=17.817,18.538;all P＜0.001),and was negatively correlated with OS in gastric cancer patients(r=-0.728,P＜0.001).In addition,qRT-PCR and in vitro co-culture results showed that the angiogenesis capacity of HUVEC cells cultured with miR-204 overexpression conditioned medium was significantly decreased(P＜0.001),while the angiogenesis capacity of HUVEC cells cultured with miR-204 knockdown conditioned medium was significantly increased(P＜0.01).Finally,western blot results showed that miR-204 overexpression significantly inhibited ZEB1 expression and induced the inactivation of VEGF/VEGFR signaling pathway in HGC-27 cells(P＜0.001).Conclusion Upregulation of miR-204 expression could inhibit angiogenesis of gastric cancer by inducing the inactivation of ZEB 1/VEGF/VEGFR signaling pathway.
Gastric cancermiR-204Zinc finger E-box-binding protein 1
NETL
NSTL
万方数据
