Guo Dong-lin, Zhou Hong, Wu Ying, Zhou Fang, Zhang Xian-mei, Xu Guo-ying, Wen Hai-ping
School of Basic Medical Sciences and Laboratory Medicine, Jiangsu University, Zhenjiang, China.
Zhonghua Zhong Liu Za Zhi. 2011 Sep;33(9):649-53.
To explore the roles of NF-κB in factor VIIa-induced proliferation and migration of a colon cancer cell line (SW620) in vitro and its possible mechanism.
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.
Factor VIIa down-regulated the IκB-α level in SW620 cells and increased the intranuclear level of NF-κB. Those effects of factor VIIa were blocked by anti-TF or anti-PAR2 antibodies. The effects of factor VIIa on proliferation and migration of SW620 cells, expression of IL-8, TF as well as caspase-7, were interfered by PDTC (the inhibitor of NF-κB).
TF/VIIa complex activates NF-κB pathway via PAR2, thereby up-regulates IL-8 and down-regulates caspase-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.
探讨核因子κB(NF-κB)在凝血因子VIIa诱导的结肠癌细胞系(SW620)体外增殖和迁移中的作用及其可能机制。
采用蛋白质免疫印迹法(Western-blotting)和实时荧光定量聚合酶链反应(real-time PCR)检测凝血因子VIIa、NF-κB抑制剂(PDTC)等处理后SW620细胞中NF-κB(p65)、NF-κB抑制蛋白(IκB-α)、半胱天冬酶-7(caspase-7)、白细胞介素8(IL-8)和组织因子(TF)的表达水平。分别通过流式细胞术和Transwell实验分析细胞的增殖和迁移情况。
凝血因子VIIa可下调SW620细胞中IκB-α水平,增加NF-κB的核内水平。抗TF或抗蛋白酶激活受体2(PAR2)抗体可阻断凝血因子VIIa的上述作用。NF-κB抑制剂(PDTC)可干扰凝血因子VIIa对SW620细胞增殖、迁移以及IL-8、TF和caspase-7表达的影响。
TF/VIIa复合物通过PAR2激活NF-κB信号通路,从而上调SW620细胞中IL-8表达,下调caspase-7表达,最终促进结肠癌细胞的增殖和迁移。此外,TF/VIIa/PAR2/NF-κB信号通路还上调TF表达,从而形成TF/VIIa/PAR2/NF-κB/TF正反馈环路。