核因子κB在凝血因子Ⅶa促进结肠癌SW620细胞增殖迁移中的作用  

Role of NF-κB in factor Ⅶa-induced proliferation and migration of colon cancer cell line SW620 cells

作　　者：郭东琳[1] 周红[1] 吴莺[2] 周芳[1] 张先梅[1] 许国莹[1] 文海平[1] 

机构地区：[1]江苏大学基础医学与医学技术学院,镇江212013 [2]江苏大学附属医院消化内科

出　　处：《中华肿瘤杂志》2011年第9期649-653,共5页Chinese Journal of Oncology

基　　金：基金项目：江苏省自然科学基金（BK2010336）

摘　　要：目的探讨核因子KB（NF—κB）在促进结肠癌SW620细胞增殖迁移过程中的作用及其可能的机制。方法以凝血因子VIIa、NF—κB抑制剂二硫代氨基甲酸吡咯烷（PDTC）等处理结肠癌细胞株SW620，采用Westernblot法检测细胞核NF—κB（p65）、细胞浆NF-κB抑制蛋白（IκB-α）和凋亡蛋白半胱氨酸蛋白酶7（caspase-7）的蛋白表达变化；采用流式细胞术检测SW620细胞的细胞周期变化；采用Transwell法测定SW620细胞的迁移能力；采用实时定量聚合酶链反应（PCR）检测白细胞介素8（IL-8）和组织因子（TF）mRNA的表达水平。结果凝血因子Ⅶa能够显著下调细胞浆中IκB-α的表达水平，并使细胞核内NF—κB的表达水平升高，单克隆抗TF和抗蛋白酶激活受体2（PAR2）抗体能够抑制凝血因子Ⅶa的这一作用。PDTC能够明显干预凝血因子Ⅶa对SW620细胞增殖、迁移的促进作用。PDTC能够明显干预凝血因子Ⅶa对SW620细胞中TF、IL-8mRNA表达的促进作用和对caspase-7蛋白表达的下调作用。结论凝血因子Ⅶa与细胞表面TF结合形成TF／Ⅶa复合物，活化受体PAR2，经NF—κB通路上调IL-8、下调caspase-7的表达，促进SW620细胞的增殖与迁移。TF／Ⅶa／／PAR2／NF—κB通路还可进一步上调TF的表达，从而形成TF／Ⅶa／PAR2／NF—κB／TF正反馈通路。Objective To explore the roles of NF-κB in factor VII a-induced proliferation and migration of a colon cancer cell line （SW620） in vitro and its possible mechanism. Methods The expression levels of NF-κB （ p65 ）, inhibitory protein of NF-κB （ IκB-α）, caspase-7, interleukin 8 （IL-8） and tissue factor （TF） in SW620 cells treated with factor Ⅶa, PDTC （an inhibitor of NF-κB） and other factors were measured by Western-blotting and real-time PCR. Proliferation and migration of the cells were analyzed by flow cytometry and Transwell assay, respectively. Results Factor Ⅶa down-regulated the IKB- ct level in SW620 cells and increased the intranuclear level of NF-KB. Those effects of factor Ⅶ a were blocked by anti-TF or anti-PAR2 antibodies. The effects of factor Ⅶ a on proliferation and migration of SW620 cells, expression of IL-8, TF as well as easpase-7, were interfered by PDTC （ the inhibitor of NF- κB）. Conclusions TF/Ⅶa complex activates NF-κB pathway via PAR2, thereby up-regulates IL-8 and down-regulates easpase-7 expression in SW620 cells, finally promotes proliferation and migration of colon cancer cells. In addition, TF/Ⅶa/PAR2/NF-κB pathway also upregulates TF expression, thus to create a positive feedback loop of TF/Ⅶa/PAR2/NF-κB/TF.

关 键 词：SW620细胞系 结肠肿瘤 凝血因子Ⅶa 蛋白酶激活受体2 核因子ΚB 

分 类 号：R735.35[医药卫生—肿瘤]