Zhang Hao, Liu Xiao-Fang, Li Yu-Ji, Zhou Jian-Ping, Kong Fan-Min, Dong Ming
Department of Surgery, The First Affiliated Hospital of China Medical University, Shenyang 110001, China.
Zhonghua Zhong Liu Za Zhi. 2007 Dec;29(12):909-12.
To determine the effect of EGF on the invasiveness of pancreatic cancer cells and its related regulatory mechanism.
The effects of EGF on the proliferation, adhesion and invasion of pancreatic cancer cells were detected by WST-1 proliferation assay, adhesion assay and invasive assay. The expression of uPA was assayed by Western blot and RT-PCR. The activity of NF-kappaB was examined by EMSA.
EGF significantly increased the invasiveness of pancreatic cancer cells but did not affect cell proliferation or adhesion. Increased invasiveness was associated with the induction of uPA at both mRNA and protein levels. Furthermore, EGF stimulated the NF-kappaB binding activity, and pretreatment of cells with a NF-kappaB inhibitor, pyrrolidine dithiocarbamate, markedly attenuated EGF-induced NF-kappaB activation. Subsequently, the EGF-induced uPA expression and invasiveness were also inhibited by NF-kappaB inhibitor.
Our findings indicated that NF-kappaB-mediated up-regulation of uPA expression is responsible for EGF-induced invasiveness in pancreatic cancer cells, and implicate that such anti-NF-kappaB therapy with NF-kappaB inhibitors may contribute to the reduction of invasiveness of pancreatic cancer.
确定表皮生长因子（EGF）对胰腺癌细胞侵袭性的影响及其相关调控机制。
采用WST - 1增殖试验、黏附试验和侵袭试验检测EGF对胰腺癌细胞增殖、黏附和侵袭的影响。通过蛋白质免疫印迹法（Western blot）和逆转录 - 聚合酶链反应（RT - PCR）检测尿激酶型纤溶酶原激活剂（uPA）的表达。采用电泳迁移率变动分析（EMSA）检测核因子κB（NF - κB）的活性。
EGF显著增加胰腺癌细胞的侵袭性，但不影响细胞增殖或黏附。侵袭性增加与uPA在mRNA和蛋白质水平的诱导有关。此外，EGF刺激NF - κB结合活性，用NF - κB抑制剂吡咯烷二硫代氨基甲酸盐预处理细胞可显著减弱EGF诱导的NF - κB激活。随后，NF - κB抑制剂也抑制了EGF诱导的uPA表达和侵袭性。
我们的研究结果表明，NF - κB介导的uPA表达上调是EGF诱导胰腺癌细胞侵袭性的原因，提示用NF - κB抑制剂进行这种抗NF - κB治疗可能有助于降低胰腺癌的侵袭性。
