水蛭素是凝血酶的特异性抑制剂之一,已被证实具有强大的生物活性,包括抑制肿瘤.然而,目前尚不清楚水蛭素和蛋白酶激活受体1(PAR-1)在弥漫性大B细胞淋巴瘤(DLBCL)中的作用和机制。通过RT-qPCR和IHC检测DLBCL组织和细胞中PAR-1的表达。转染sh-NC,sh-PAR-1或pcDNA3.1-PAR-1在DLBCL细胞或通过添加凝血酶处理的DLBCL细胞中,Vorapaxar,重组水蛭素(RH),或Na2S2O4并与EA共培养。hy926.并构建DLBCL小鼠观察肿瘤生长情况。RT-qPCR检测相关基因的表达,蛋白质印迹,IHC,和免疫荧光,用低氧探针-1试剂盒测量细胞缺氧,并估计细胞炎症因子,扩散,迁移,入侵,和细胞凋亡通过ELISA,CCK-8,流式细胞术,伤口愈合和Transwell.使用免疫共沉淀和下拉测量来验证该关系。PAR-1在DLBCL组织和细胞中高表达,特别是在SUDHL2。Na2S2O4诱导SUDHL2缺氧,PAR-1不影响凝血酶激活的缺氧。PAR-1可以促进SUDHL2的增殖,迁移,和入侵,与细胞缺氧无关.PAR-1促进增殖,迁移,和EA的血管生成。hy926或SUDHL2通过上调血管内皮生长因子(VEGF)。RH抑制肿瘤生长,细胞增殖,和移民,促进DLBCL的凋亡,并通过下调PAR-1-VEGF抑制血管生成。RH抑制增殖,迁移,和通过下调PAR-1-VEGF的DLBCL细胞的血管生成。
Hirudin is one of the specific inhibitors of thrombin, which has been confirmed to have strong bioactivities, including inhibiting tumors. However, the function and mechanism of hirudin and protease-activated receptor 1 (PAR-1) in diffuse large B-cell lymphoma (DLBCL) have not been clear. Detecting the expression PAR-1 in DLBCL tissues and cells by RT-qPCR and IHC. Transfected sh-NC, sh-PAR-1, or pcDNA3.1-PAR-1 in DLBCL cells or processed DLBCL cells through added thrombin, Vorapaxar, Recombinant hirudin (RH), or Na2S2O4 and co-culture with EA.hy926. And built DLBCL mice observed tumor growth. Detecting the expression of related genes by RT-qPCR, Western blot, IHC, and immunofluorescence, measured the cellular hypoxia with Hypoxyprobe-1 Kit, and estimated the cell inflammatory factors, proliferation, migration, invasion, and apoptosis by ELISA, CCK-8, flow cytometry, wound-healing and Transwell. Co-immunoprecipitation and pull-down measurement were used to verify the relationship. PAR-1 was highly expressed in DLBCL tissues and cells, especially in SUDHL2. Na2S2O4 induced SUDHL2 hypoxia, and PAR-1 did not influence thrombin-activated hypoxia. PAR-1 could promote SUDHL2 proliferation, migration, and invasion, and it was unrelated to cellular hypoxia. PAR-1 promoted proliferation, migration, and angiogenesis of EA.hy926 or SUDHL2 through up-regulation vascular endothelial growth factor (VEGF). RH inhibited tumor growth, cell proliferation, and migration, promoted apoptosis of DLBCL, and inhibited angiogenesis by down-regulating PAR-1-VEGF. RH inhibits proliferation, migration, and angiogenesis of DLBCL cells by down-regulating PAR-1-VEGF.