Involvement of PI3K and ERK1/2 pathways in hepatocyte growth factor-induced cholangiocarcinoma cell invasion
Involvement of PI3K and ERK1/2 pathways in hepatocyte growth factor-induced cholangiocarcinoma cell invasion
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DOI：
10.3748/wjg.v16.i6.713
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发表时间：
2010-02-14
影响因子：
4.3
通讯作者：
Suthiphongchai, Tuangporn
中科院分区：
文献类型：
--
作者：
Menakongka, Apaporn;Suthiphongchai, Tuangporn
目的：为探讨肝细胞生长因子（hepatocyte growth factor，HGF）在胆管癌（cholangiocarcinoma，CCA）细胞侵袭中的作用及其机制，采用Transwell体外侵袭实验，观察HGF对人CCA细胞株HuCCA-1和KKU-M213侵袭和运动能力的影响。通过蛋白质印迹法测定目的蛋白质及其磷酸化形式的水平。通过免疫荧光染色和共聚焦显微镜观察分析E-钙粘蛋白的定位。结果：两种CCA细胞系表达的Met水平均高于H69永生化胆管细胞系。HGF诱导细胞系的侵袭性和运动性，并改变E-钙粘蛋白从膜到细胞质的定位，但不影响分泌的基质金属蛋白酶（MMP）-2，MMP-9和尿激酶纤溶酶原激活剂，参与细胞侵袭的关键基质降解酶的水平。同时，HGF刺激Akt和细胞外信号调节激酶（ERK）1/2磷酸化，但在两种细胞系中的动力学特征略有不同。用磷脂酰肌醇3-激酶（PI 3 K）抑制剂LY 294002抑制PI 3 K/Akt信号通路可显著抑制HGF诱导的两种CCA细胞系的侵袭，用U 0126抑制ERK信号通路可抑制HGF诱导的KKU-M213细胞系的侵袭，但对HuCCA-1细胞有中等程度的影响。这些数据表明，肝细胞生长因子促进CCA细胞的侵袭性，通过异位定位的E-钙粘蛋白和诱导细胞运动的不同信号转导途径依赖于细胞系类型。(C)2010年百世登。All rights reserved.
AIM: To investigate the role of hepatocyte growth factor (HGF) in cholangiocarcinoma (CCA) cell invasiveness and the mechanisms underlying such cellular responses.METHODS: Effects of HGF on cell invasion and motility were investigated in two human CCA cell lines, HuCCA-1 and KKU-M213, using Transwell in vitro assay. Levels of proteins of interest and their phosphorylated forms were determined by Western blotting. Localization of E-cadherin was analyzed by immunofluorescence staining and visualized under confocal microscope. Activities of matrix degrading enzymes were determined by zymography.RESULTS: Both CCA cell lines expressed higher Met levels than the H69 immortalized cholangiocyte cell line. HGF induced invasion and motility of the cell lines and altered E-cadherin from membrane to cytoplasm localization, but did not affect the levels of secreted matrix metalloproteinase (MMP)-2, MMP-9 and urokinase plasminogen activator, key matrix degrading enzymes involved in cell invasion. Concomitantly, HGF stimulated Akt and extracellular signal-regulated kinase (ERK)1/2 phosphorylation but with slightly different kinetic profiles in the two cell lines. Inhibition of the phosphoinositide 3-kinase (PI3K)/Akt pathway by the PI3K inhibitor, LY294002, markedly suppressed HGF-stimulated invasion of both CCA cell lines, and inhibition of the ERK pathway by U0126 suppressed HGF-induced invasion of the KKU-M213 cell line but had a moderate effect on HuCCA-1 cells.CONCLUSION: These data indicate that HGF promotes CCA cell invasiveness through dys-localization of E-cadherin and induction of cell motility by distinct signaling pathways depending on cell line type. (C) 2010 Baishideng. All rights reserved.