Mechanisms of invasion induced by hepatocyte growth factor in gallbladder carcinoma cell lines

肝细胞生长因子诱导胆囊癌细胞侵袭的机制

• 批准号: 07671400
• 负责人: SHIMURA Hideo
• 金额: $ 1.47万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671400/
• 关键词: gallbladder cancer cells; cancer invasion; HGF; c-met; proteolytic enzymes; fibroblasts; cancer-mesenchymal interaction; 胆嚢癌細胞株; 浸潤; タンパク分解酵素

Human gallbladder cancers are highly malignant and its prognosis is poor depending on extend of surround tissue invasion. We examined in vitro the invasive activity of four gallbladder cancer cell lines (GB-d1, GB-h3, GB-d2 and FU-GBC-1) in the absence or presence of hepatocyte growth factor (HGF). In type 1 collagen gel culture, HGF stimulated cell proliferation and induced invasive phenotype of arborizing or individual tubule structures in GB-d1, GB-h3 and GB-d2. In Matrigel invasion assay, invasion was also induced in these three cell lines by HGF but not in FU-GBC-1. Cellular motility was however stimulated by HGF in all of the four cell lines at various extent. HGF was produced from primary cultured normal fibroblasts of human gallbladders but not from the gallgaldder cancer cell lines. Zymography for proteolytic enzymes demonstrated high levels of type IV collagenase and urokinase-type plasminogen activator (u-PA) activity in GB-d1, GB-h3 and GB-d2 even in the absence of HGF.In the presence of HGF,the 72K type IV collagenase (MMP-2) activity of GB-h3 and u-PA activities of GB-d1, GB-h3 and GB-d2 were enhanced. In contrast, MMPs and PAs activities of FU-GBC-1 were faint irrespective of HGF addition. Western blot analysis demonstrated higher levels of 190K c-met product (HGF receptor) in GB-d1, GB-h3 and GB-d2 than FU-GBC-1. These results suggest that in addition to the importance of proteolytic potent, the cellular motility induced via HGF/HGF-receptor system is essential for the invasive progression of the gallbladder carcinoma cells.

胆囊癌恶性程度高，其预后取决于周围组织的侵袭程度。我们检测了四种胆囊癌细胞系(GB-D1、GB-H3、GB-D2和FU-GBC-1)在无或有肝细胞生长因子(HGF)作用下的体外侵袭活性。在1型胶原凝胶培养中，HGF刺激细胞增殖，并诱导GB-D1、GB-H3和GB-D2的树枝状或单个小管结构的侵袭表型。在Matrigel侵袭实验中，HGF也能诱导这三种细胞的侵袭，而FU-GBC-1则不能。HGF对四种细胞系的细胞运动均有不同程度的刺激作用。肝细胞生长因子是从原代培养的人胆囊正常成纤维细胞产生的，而不是从胆囊癌细胞系产生的。蛋白水解酶的酶谱分析显示，即使在没有HGF的情况下，GB-D1、GB-H3和GB-D2中也有较高水平的IV型胶原酶和尿激酶型纤溶酶原激活物(u-PA)活性。而FU-GBC-1的MMPs和PAS活性较弱，与HGF的添加无关。Western印迹分析显示，Gb-d1、Gb-h3和Gb-D2的190K c-Met产物(HGF受体)的表达水平高于FU-GBC-1。这些结果表明，除了蛋白水解力的重要性外，通过HGF/HGF受体系统诱导的细胞运动对胆囊癌细胞的侵袭发展也是必不可少的。

项目成果

Shimura H: "Induction of invasive growth in a gallbladder cancer cell line by hepatocyte growth factor in vitro." Jpn Cancer Res. 86. 662-669 (1995)

Shimura H：“体外肝细胞生长因子诱导胆囊癌细胞系的侵袭性生长。”

Aoki Y: "Role of hepatocyte growth factor derived from human gallbladder fibroblasts in the invasion model of gallbladder cancer." J Jpn Res Soc Gastroenterol Carcinog. 7. 255-58 (1995)

Aoki Y：“源自人胆囊成纤维细胞的肝细胞生长因子在胆囊癌侵袭模型中的作用。”

Aoki Y,Shimura H,Li Hong, Date K,Tanaka M: "Role of hepatocyte growth factor derived from human gallbladder fibroblasts in the invasion model of gallbladder cancer." J Jpn Res Soc Gastroenterol Carcinog. 7. 255-58 (1995)

Aoki Y，Shimura H，Li Hong，Date K，Tanaka M：“人胆囊成纤维细胞衍生的肝细胞生长因子在胆囊癌侵袭模型中的作用。”

Shimura H,Date K,Matsumoto K,Nakamura T,Tanaka M: "Induction of invasive growth in a gallbladder cancer cell line by hepatocyte growth factor in vitro." Jpn Cancer Res. 86. 662-9 (1995)

Shimura H、Date K、Matsumoto K、Nakamura T、Tanaka M：“体外肝细胞生长因子诱导胆囊癌细胞系的侵袭性生长。”

Matsumoto K: "Acquisition of invasive phenotype in galbladder cancer cells via mutual interaction of stromal fibroblasts and cancer cells as mediated by hepatocyte growth factor." Jpn Cancer Res. 87. 702-710 (1996)

Matsumoto K：“通过肝细胞生长因子介导的基质成纤维细胞和癌细胞的相互作用，获得胆囊癌细胞的侵袭性表型。”