膀胱癌组织中TIPE通过调节VEGFR2表达促进膀胱癌细胞增殖、迁移和血管生成

TIPE in bladder cancer tissues promotes cell proliferation,migration and angiogenesis of bladder cancer by regulating the expression of VEGFR2

目的:探究膀胱癌组织中TIPE通过调节血管内皮生长因子受体2(VEGFR2)表达促进膀胱癌细胞增殖、迁移和血管生成的机制。方法:收集膀胱癌组织及其对应的癌旁组织各34例,以人膀胱上皮永生化细胞SV-HUC-1、人膀胱癌细胞株UMUC3和人脐静脉内皮细胞(HUVECs)为体外研究对象,免疫组织化学(IHC)检测组织中TIPE表达;RT-PCR检测细胞中TIPE mRNA表达;Western blot检测组织和细胞中TIPE、PDK1、p-PDK1和VEGFR2蛋白表达;CCK-8检测UMUC3细胞增殖能力;Transwell实验检测UMUC3细胞迁移能力;血管形成实验检测HUVECs血管形成能力。结果:与癌旁组织相比,膀胱癌组织中TIPE IHC评分和蛋白表达明显上调(P<0.05)。与SV-HUC-1组相比,UMUC3组中TIPE mRNA和蛋白表达水平明显上调(P<0.05)。与siRNA-NC组相比,siRNA-TIPE组UMUC3细胞增殖、迁移能力、HUVECs的管道交叉数明显降低(P<0.05),TIPE、p-PDK1和VEGFR2蛋白表达水平明显降低(P<0.05)。PDK1抑制剂GSK2334470(简称GSK)的使用浓度为4μmol/L。与Vector组相比,TIPE组UMUC3细胞TIPE、p-PDK1和VEGFR2蛋白表达水平及细胞增殖、迁移能力和HUVECs的管道交叉数明显升高(P<0.05),Vector+GSK组UMUC3细胞p-PDK1和VEGFR2蛋白表达水平及细胞增殖、迁移能力和HUVECs的管道交叉数明显降低(P<0.05),GSK可逆转TIPE对UMUC3和HUVECs细胞的作用。结论:TIPE通过磷酸化PDK1上调VEGFR2表达促进膀胱癌细胞增殖、迁移和血管生成。

Objective:To explore the mechanism of TIPE in bladder cancer tissues promotes cell proliferation,migration and angiogenesis of bladder cancer by regulating the expression of vascular endothelial growth factor receptor 2(VEGFR2).Methods:Thirty-four cases of bladder cancer tissues and its corresponding adjacent tissues were collected.Human bladder epithelial immortalized cells SV-HUC-1,human bladder cancer cell line UMUC3 and human umbilical vein endothelial cells(HUVECs)were used as the in vitro study objects.The immunohistochemistry(IHC)method was used to detect TIPE expression in tissues.RT-PCR was used to detect TIPE mRNA expression in cells.Western blot was used to detect the protein expression of TIPE,PDK1,p-PDK1 and VEGFR2 in tissues and cells.CCK-8 was used to detect UMUC3 cells proliferation ability.The Transwell test was used to detect the migration ability of UMUC3 cells.The angiogenesis test was used to detect the vascularization ability of HUVECs.Results:Compared with adjacent tissues,TIPE IHC score and protein expression in bladder cancer tissues were significantly up-regulated(P<0.05).Compared with the SV-HUC-1 group,the mRNA and protein expression levels of TIPE were significantly up-regulated in the UMUC3 group(P<0.05).Compared with the siRNA-NC group,the UMUC3 cells proliferation,migration ability and the crossing number of HUVECs pipeline in the siRNA-TIPE group were significantly reduced(P<0.05),and the protein expression levels of TIPE,p-PDK1 and VEGFR2 were significantly reduced(P<0.05).The concentration of PDK1 inhibitor GSK2334470(GSK for short)was 4μmol/L.Compared with the Vector group,the protein expression levels of TIPE,p-PDK1 and VEGFR2 of UMUC3 cells,UMUC3 cells proliferation and migration ability,the crossing number of HUVECs pipeline in the TIPE group were significantly increased(P<0.05),meanwhile,the protein expression levels of p-PDK1 and VEGFR2 of UMUC3 cells,UMUC3cells proliferation and migration ability,the crossing number of HUVECs pipeline in Vector+GSK group were signifi-cantly reduced(P<0.05).GSK can reverse the effect of TIPE on UMUC3 and HUVECs cells.Conclusion:TIPE up-regulates the expression of VEGFR2 through phosphorylation of PDK1 to promote bladder cancer cell proliferation,mi-gration and angiogenesis.