Tumor-derived trypsin enhances proliferation of intrahepatic cholangiocarcinoma cells by activating protease-activated receptor-2
Tumor-derived trypsin enhances proliferation of intrahepatic cholangiocarcinoma cells by activating protease-activated receptor-2
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DOI：
10.3892/ijo_00000555
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发表时间：
2010-04-01
影响因子：
5.2
通讯作者：
Harada, Shin-Ichi
中科院分区：
文献类型：
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作者：
Nakanuma, Shin-ichi;Tajima, Hidehiro;Harada, Shin-Ichi
在原发性恶性肝肿瘤中。70%的肝内胆管细胞癌(ICC)标本中存在胰蛋白酶原免疫反应，而肝细胞癌(HCC)标本中无胰蛋白酶原免疫反应。我们认为，胰酶原的分泌是ICC细胞和肝癌细胞生物学行为的关键区别。本研究的目的是利用细胞系和手术标本研究肿瘤源性胰酶的分泌及其特异性受体-2(PAR-2)在ICC中的表达。在4株ICC细胞中有3株表达胰蛋白酶原-1mRNA，而在3株肝癌细胞株中均未见表达。Western印迹分析在无血清条件培养液中检测到胰蛋白酶原-1，该细胞系的一株ICC细胞表达该基因。明胶酶谱显示，在相同的条件介质中，胰蛋白酶原的活化形式胰蛋白酶具有明胶溶解活性。在ICC细胞系中可观察到PAR-2mRNA和蛋白的表达。浓度低至10 nM的胰酶可促进ICC细胞的增殖，100 nM时达高峰。丝氨酸蛋白酶抑制剂甲磺酸加贝酯可抑制胰酶的作用。免疫组织化学方法检测到%的胆囊癌组织中有PAR-2表达。此外，52%的ICC间质成纤维细胞表达PAR-2。这些结果表明，胰蛋白酶原-1促进了ICC细胞和肿瘤相关成纤维细胞的生长。
In primary malignant liver tumors. trypsinogen-immunoreactivity was present in 70% of intrahepatic cholangiocarcinoma (ICC) specimen, but absent in hepatocellular carcinoma (HCC) specimens. We suggest the secretion of trypsinogen to be a key difference in biological behavior between ICC and HCC cells. The purpose of this study was to investigate the secretion of tumor-derived trypsin and the expression of its specific receptor, protease-activated receptor-2 (PAR-2), in ICC using cell lines and surgical specimens. The expression of trypsinogen-1 mRNA was observed in three of four ICC cell lines, but none of three HCC cell lines. Western blot analysis detected trypsinogen-1 in serum-free conditioned medium from one of the ICC cell lines positive for the mRNA. Gelatin zymography revealed a gelatinolytic activity for trypsin, the activated form of trypsinogen, in the same conditioned medium. PAR-2 mRNA and protein were observed in ICC cell lines. The proliferative activity of ICC cells was increased by concentrations of trypsin as low as 10 nM, and peaked at 100 nM. The effect of trypsin was suppressed by a serine protease inhibitor, gabexate mesilate. PAR-2 expression was detected in 64% of ICC surgical specimens immunohistochemically. In addition, stroma fibroblasts expressed PAR-2 in 52% of ICC specimens. These results suggest that trypsinogen-1 contributes to the growth of ICC cells and also tumor-associated fibroblasts.