幽门螺杆菌刺激胃癌细胞株SGC-7901分泌白细胞介素-8过程中核因子κB的作用  

The role of nuclear factor kappa B in secretion of interleukin-8 by gastric cancer cell line SGC 7901 induced by helicobacter pylori

作　　者：史彤[1] 刘文忠[1] 高飞[2] 萧树东[1] 

机构地区：[1]上海第二医科大学附属仁济医院上海市消化疾病研究所,200001 [2]上海第二医科大学细胞生物学教研室

出　　处：《中华医学杂志》2003年第2期133-136,共4页National Medical Journal of China

摘　　要：目的　探讨胃上皮细胞在幽门螺杆菌 (Hp)刺激下分泌白细胞介素 8(IL 8)的过程中 ,核因子 κB(NF κB)的作用。方法　电穿孔法将I κBαM(NF κB抑制蛋白 )基因转染入SGC 790 1中 ,β半乳糖苷酶分析转染效率 ,Western印迹分析I κB基因的表达。将不同浓度的Hp活菌、灭活菌及液体培养上清分别与稳定转染I κB的胃癌细胞株SGC 790 1及其空白质粒对照共同孵育 ,凝胶电泳迁移率变迁分析 (EMSA)和NF κB荧光素酶报告基因分析检测不同时间细胞内NF κB的激活 ,ELISA法检测不同时间细胞上清中IL 8的水平。结果　得到了稳定表达I κB的SGC 790 1细胞 ,命名为SGC790 1 I κB细胞 ,其空白质粒对照命名为SGC790 1 neo。Hp活菌刺激的SGC790 1 neo在 2h和 4h时可检测到明显的NF κB激活 ,灭活菌和液体培养上清刺激下无激活 ;Hp活菌、灭活菌和培养上清刺激下SGC 790 1 I κB内均无NF κB激活。 4h时 ,2× 10 7Hp/ml的Hp活菌刺激下即可检测到SGC790 1 neo内明显的IL 8分泌 ,并有时间 剂量依赖效应 ;灭活菌和液体培养上清无此作用。Hp活菌、灭活菌和液体培养上清均不能刺激SGC790 1 I κB产生IL 8。结论　Hp诱导胃上皮细胞分泌IL 8依赖于NF κB的激活。Objective To investigate the role of nuclear factor κB (NF-κB) in the gastric inflammation induced by Helicobacter pylori(H. pylori). Methods SGC-7901 was transfected with IκB(NF-κB inhibitor gene) by electroporation, βlacZ activity assay was used to examine transfected efficacy. Expression of IκB was assessed by Western-blot. Different concentration of live and heat-killed Hp (ATCC 43504) and supernatant of liquid culture were cocultured with SGC7901- IκB and its negative control SGC7901- neo. Activation of intracellular NF-κB was examined by electrophoretic mobility shift analyses(EMSA) and luciferase report gene assay at different time point. IL-8 levels were measured by ELISA at different time point. Results IL-8 release was evident 4 hours after infection of SGC-7901-neo with H.pylori, and this effect was dose-time dependent. SGC-7901-IκB in which NF-κB has not been activated could not secret IL-8 after infection with H. pylori. Conclusion Secretion of IL-8 by gastric epithelial cell upon H pylori infection is dependent on activation of NF-κB.

关 键 词：幽门螺杆菌 胃癌 细胞株 SGC-7901 白细胞介素-8 核因子ΚB 

分 类 号：R735.2[医药卫生—肿瘤]