硼替佐米对人胃癌SGC-7901细胞uPA、NF-κB表达的影响及与其侵袭力关系

Effect of bortezomib on the expression of NF-κB and UPA as well as cell invasiveness in human SGC-7901 gastric cancer cells

目的硼替佐米对人胃癌SGC-7901细胞uPA、NF-κB表达的影响及与其侵袭力关系。方法 Brdu ELISA法测细胞增殖活性;Boyden小室培养测细胞的迁移率;Western blot测细胞uPA、NF-κB的蛋白水平;细胞免疫化学测NF-κB细胞表达及细胞定位。结果 (1)与对照组(无血清培养基组)相比,10%FCS组细胞增殖活性与迁移率明显增高(P<0.05);与10%FCS组相比,硼替佐米呈浓度依赖性抑制人胃癌SGC-7901细胞增殖及迁移,硼替佐米浓度为(4μg.L-1),人胃癌SGC-7901细胞增殖活性与迁移率均明显降低(P<0.05);与PDTC组(10μmol.L-1)相比,两者无明显差别;(2)与10%FCS组相比,硼替佐米呈浓度依赖性抑制胃癌SGC-7901细胞uPA、NF-κB蛋白表达,硼替佐米浓度为(4μg.L-1)时uPA、NF-κB蛋白表达均明显降低(P<0.05);与NF-κB特异性抑制剂PDTC组相比,两组uPA、NF-κB表达均无明显差别;硼替佐米浓度为(4μg.L-1)能明显降低NF-κB核蛋白含量,与PDTC组相比,两者无显著差异(P>0.05);(4)细胞免疫化学结果示:硼替佐米(4μg.L-1)抑制10%FCS诱导人胃癌SGC-7901细胞NF-κB核移位。结论①硼替佐米抑制血清诱导胃癌SGC-7901细胞增殖迁移及uPA、NF-κB蛋白表达,②硼替佐米降低胃癌SGC-7901细胞侵袭力可能与其抑制NF-κB活性,降低UPA水平有关。

Aim To observe the effect of bortezomib on the expression of NF-κB and UAP as well as cell invasiveness in human SGC-7901 gastric cancer cells.Methods Proliferation activity and migration level of human SGC-7901 cells were respectively determined by Bromodeoxyuridine（BrdU） ELISA and Boyden chamber.The expression of NF-κB and uPA in human SGC-7901 cells was determined by Western blot.The expression and localization of NF-κB in human SGC-7901 cells were further revealed by immunocytochemistry.Results Compared with the control group（the serum-free medium group）,proliferation and migration of human SGC-7901 cells were significantly increased in 10% FCS group（P0.05）.Proliferation and migration of human SGC-7901 cells were inhibited by bortezomib in a concentration-dependent manner,which was significantly decreased by bortezomib with a concentration of 4 μg·L-1 compared with 10% FCS group（P0.05）,but were similar PDTC（10 μmol·L-1） group.Compared with 10% FCS group,the expression of uPA and NF-κB in human SGC-7901 cells was inhibited by bortezomib in a concentration dependent manner.The expression of uPA and NF-κB in the group of bortezomib at 4 μg·L-1 was significantly decreased when compared with the 10% FCS group（P0.05）,but were similar to PDTC group（P0.05）.The expression of NF-κB-p65 nuclear protein in the group of bortezomib at 4 μg·L-1 was significantly decreased when compared with 10% FCS group（P0.05）,but was similar to PDTC group（P0.05）.Immunocytochemistry showed that NF-κB nuclear translocation in human SGC-7901 cells induced by 10% FCS was inhibited by bortezomib with a concentration of 4 μg·L-1.Conclusions ① Bortezomib inhibits the expression of uPA and NF-κB as well as cell proliferation and migration in human SGC-7901 cells.② The inhibition of bortezomib on the invasiveness of human SGC-7901 cells may relate to its inhibition on NF-κB activity and UPA expression.