Heparin regulates colon cancer cell growth through p38 mitogen-activated protein kinase signalling
Heparin regulates colon cancer cell growth through p38 mitogen-activated protein kinase signalling
复制标题
DOI：
10.1111/j.1365-2184.2009.00649.x
复制
发表时间：
2010-02-01
影响因子：
8.5
通讯作者：
Tzanakakis, G. N.
中科院分区：
文献类型：
--
作者：
Chatzinikolaou, G.;Nikitovic, D.;Tzanakakis, G. N.
目的：肝素是一种细胞外刺激物，能够激活主要的细胞信号通路。因此，我们探讨了肝素刺激HT29、SW1116和HCT116细胞生长的可能机制。材料和方法：通过特异性MAPK级联抑制剂、Western印迹分析、实时定量聚合酶链式反应和FACS细胞凋亡分析，检测丝裂原活化蛋白激酶(MAPK)级联通路在肝素诱导的HT29、SW1116和HCT116细胞生长中的可能参与。结果：用高度特异的p38激酶抑制剂SB203580处理后，显著(50-70%)抑制肝素诱导的结肠癌细胞生长，表明p38 MAPK信号转导参与了肝素诱导的结肠癌细胞增殖反应。这与p38 MAP上181/182苏氨酸/酪氨酸残基的磷酸化增加(最多3倍)有关。此外，肝素通过p38介导的机制抑制细胞周期蛋白依赖性激酶抑制因子p21WAF1/CIP1和p53抑癌基因和蛋白的表达，使细胞周期蛋白D1的表达增加1.8倍。另一方面，肝素对HT29、SW1116和HCT116细胞的凋亡水平没有影响。结论：本研究表明，胞外糖胺多糖肝素通过特异性激活p38丝裂原蛋白激酶刺激结肠癌细胞生长，精细调控细胞周期调控关键基因的表达。
Objectives:Heparin acts as an extracellular stimulus capable of activating major cell signalling pathways. Thus, we examined the putative mechanisms utilized by heparin to stimulate HT29, SW1116 and HCT116 colon cancer cell growth.Materials and methods:Possible participation of the mitogen-activated protein kinase (MAPK) cascade on heparin-induced HT29, SW1116 and HCT116 colon cancer cell growth was evaluated using specific MAPK cascade inhibitors, Western blot analysis, real-time quantitative PCR and FACS apoptosis analysis.Results:Treatment with a highly specific p38 kinase inhibitor, SB203580, significantly (50-70%) inhibited heparin-induced colon cancer cell growth, demonstrating that p38 MAPK signalling is involved in their heparin-induced proliferative response. This was shown to be correlated with increased (up to 3-fold) phosphorylation of 181/182 threonine/tyrosine residues on p38 MAP kinase. Furthermore, heparin inhibited cyclin-dependent kinase inhibitor p21WAF1/CIP1 and p53 tumour suppressor gene and protein expression up to 2-fold or 1.8-fold, respectively, and stimulated cyclin D1 expression up to 1.8-fold, in these cell lines through a p38-mediated mechanism. On the other hand, treatment with heparin did not appear to affect HT29, SW1116 and HCT116 cell levels of apoptosis.Conclusions:This study demonstrates that an extracellular glycosaminoglycan, heparin, finely modulates expression of genes crucial to cell cycle regulation through specific activation of p38 MAP kinase to stimulate colon cancer cell growth.