Hepatocyte growth factor induces colonic cancer cell invasiveness via enhanced motility and protease overproduction. Evidence for PI3 kinase and PKC involvement
Hepatocyte growth factor induces colonic cancer cell invasiveness via enhanced motility and protease overproduction. Evidence for PI3 kinase and PKC involvement
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DOI：
10.1093/carcin/22.7.1035
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发表时间：
2001-07-01
期刊：
影响因子：
4.7
通讯作者：
Lehy, T
中科院分区：
文献类型：
--
作者：
Kermorgant, S;Aparicio, T;Lehy, T
肿瘤向转移表型的进展主要依赖于肿瘤细胞的侵袭性，细胞迁移是这一过程中的关键步骤。在这里，我们研究肝细胞生长因子（HGF）对低侵袭性Caco-2结肠癌上皮细胞体外侵袭诱导的影响，通过Matrigel屏障进行侵袭试验。通过酶谱法、逆转录-聚合酶链反应和免疫印迹法对蛋白酶进行评估。发现Caco-2细胞表达HGF受体而不表达HGF，并分泌几种蛋白酶，即基质金属蛋白酶-1（MMP-1）、MMP-2，可能还有MMP-9和尿激酶纤溶酶原激活物（uPA）。外源性HGF促进Caco-2细胞通过人工基底膜基质的侵袭，并增加其蛋白酶的产生。在侵袭试验结束时对培养基的分析表明，抗HGF抗体抑制蛋白酶的产生与细胞侵袭平行。使用合成的一般MMP抑制剂或抗MMP或uPA的中和抗体进一步研究蛋白酶在HGF诱导的侵袭过程中的参与。所有组分均能显著抑制HGF促进的细胞侵袭。此外，PKC α/β 1和PI 3激酶的特异性抑制剂也能降低HGF促进的细胞侵袭和侵袭试验培养基中蛋白酶的表达。因此，我们的研究结果提供的证据表明，HGF激活的Caco-2细胞的侵袭性的过程涉及PI 3激酶和PKC，并从两个事件，刺激细胞运动活性和伴随的蛋白酶的过度生产，这使得细胞迁移通过降解的细胞外基质的密切关联的结果。
Tumour progression to the metastatic phenotype is mainly dependent on tumour cell invasiveness, Cell migration is a crucial step in this process. Here we investigate the effect of hepatocyte growth factor (HGF) on the induction of in vitro invasiveness of poorly aggressive Caco-2 colonic cancer epithelial cells, Invasion assays through a Matrigel barrier were performed. Proteases were assessed by zymography, reverse transcription-polymerase chain reaction and immunoblotting. Caco-2 cells were found to express HGF receptor but not HGF and to secrete several proteases, namely matrix metalloproteinase-1 (MMP-1), MMP-2, possibly MMP-9 and urokinase plasminogen activator (uPA), Exogenous HGF promoted invasiveness of Caco-2 cells through an artificial basement membrane matrix and enhanced their production of proteases, In addition, analyses of media at the end of invasion assays indicated that anti-HGF antibody inhibited protease production in parallel with cell invasion, The involvement of proteases in the HGF-induced invasion process was further investigated using either a synthetic general MMP inhibitor or neutralizing antibodies against MMPs or uPA. All components significantly inhibited HGF-promoted cell invasion, Moreover, specific inhibitors of PKC alpha/beta1 and PI3 kinase also decreased both HGF-promoted cell invasion and protease expression in invasion assay media. Thus, our findings provide evidence that the process of HGF-activated invasiveness of Caco-2 cells involves PI3 kinase and PKC and results from close association of two events, stimulation of cell motile activity and concomitant overproduction of proteases, which permits cell migration through a degraded extracellular matrix.