Kenny Susan, Duval Cedric, Sammut Stephen J, Steele Islay, Pritchard D Mark, Atherton John C, Argent Richard H, Dimaline Rod, Dockray Graham J, Varro Andrea
Physiological Laboratory, School of Biomedical Sciences, University of Liverpool, Crown St., Liverpool L69 3BX, UK.
Am J Physiol Gastrointest Liver Physiol. 2008 Sep;295(3):G431-41. doi: 10.1152/ajpgi.90283.2008. Epub 2008 Jul 3.
The gastric pathogen Helicobacter pylori (H. pylori) is linked to peptic ulcer and gastric cancer, but the relevant pathophysiological mechanisms are unclear. We now report that H. pylori stimulates the expression of plasminogen activator inhibitor (PAI)-1, urokinase plasminogen activator (uPA), and its receptor (uPAR) in gastric epithelial cells and the consequences for epithelial cell proliferation. Real-time PCR of biopsies from gastric corpus, but not antrum, showed significantly increased PAI-1, uPA, and uPAR in H. pylori-positive patients. Transfection of primary human gastric epithelial cells with uPA, PAI-1, or uPAR promoters in luciferase reporter constructs revealed expression of all three in H+/K+ATPase- and vesicular monoamine transporter 2-expressing cells; uPA was also expressed in pepsinogen- and uPAR-containing trefoil peptide-1-expressing cells. In each case expression was increased in response to H. pylori and for uPA, but not PAI-1 or uPAR, required the virulence factor CagE. H. pylori also stimulated soluble and cell surface-bound uPA activity, and both were further increased by PAI-1 knockdown, consistent with PAI-1 inhibition of endogenous uPA. H. pylori stimulated epithelial cell proliferation, which was inhibited by uPA immunoneutralization and uPAR knockdown; exogenous uPA also stimulated proliferation that was further increased after PAI-1 knockdown. The proliferative effects of uPA were inhibited by immunoneutralization of the EGF receptor and of heparin-binding EGF (HB-EGF) by the mutant diphtheria toxin CRM197 and an EGF receptor tyrosine kinase inhibitor. H. pylori induction of uPA therefore leads to epithelial proliferation through activation of HB-EGF and is normally inhibited by concomitant induction of PAI-1; treatments directed at inhibition of uPA may slow the progression to gastric cancer.
胃病原体幽门螺杆菌(H. pylori)与消化性溃疡和胃癌有关,但相关的病理生理机制尚不清楚。我们现在报告,幽门螺杆菌刺激胃上皮细胞中纤溶酶原激活物抑制剂(PAI)-1、尿激酶型纤溶酶原激活物(uPA)及其受体(uPAR)的表达以及对上皮细胞增殖的影响。来自胃体而非胃窦活检组织的实时PCR显示,幽门螺杆菌阳性患者的PAI-1、uPA和uPAR显著增加。用荧光素酶报告构建体中的uPA、PAI-1或uPAR启动子转染原代人胃上皮细胞,结果显示在表达H+/K+ATP酶和囊泡单胺转运体2的细胞中这三者均有表达;uPA也在表达胃蛋白酶原和含uPAR的三叶因子-1的细胞中表达。在每种情况下,表达均因幽门螺杆菌而增加,对于uPA而言,需要毒力因子CagE,但PAI-1或uPAR则不需要。幽门螺杆菌还刺激可溶性和细胞表面结合的uPA活性,并且通过敲低PAI-1两者均进一步增加,这与PAI-1对内源性uPA的抑制作用一致。幽门螺杆菌刺激上皮细胞增殖,这被uPA免疫中和及敲低uPAR所抑制;外源性uPA也刺激增殖,在敲低PAI-1后进一步增加。uPA的增殖作用被突变的白喉毒素CRM197和表皮生长因子受体酪氨酸激酶抑制剂对表皮生长因子受体和肝素结合表皮生长因子(HB-EGF)的免疫中和所抑制。因此,幽门螺杆菌诱导uPA通过激活HB-EGF导致上皮增殖,并且通常被同时诱导的PAI-1所抑制;针对抑制uPA的治疗可能会减缓向胃癌的进展。