Inhibition of Src Impairs the Growth of Met-Addicted Gastric Tumors
Inhibition of Src Impairs the Growth of Met-Addicted Gastric Tumors
复制标题
DOI：
10.1158/1078-0432.ccr-10-0106
复制
发表时间：
2010-08-01
影响因子：
11.5
通讯作者：
Trusolino, Livio
中科院分区：
文献类型：
--
作者：
Bertotti, Andrea;Bracco, Cecilia;Trusolino, Livio
目的：我们研究了抑制 Src 酪氨酸激酶（MET 癌基因的下游效应子）是否可以阻碍依赖 Met 生长和存活的胃肿瘤的恶性特性。实验设计：通过测量克隆形成存活（锚定非依赖性生长）在体外确定对 Src 抑制的敏感性，并通过建立异种移植模型在体内确定对 Src 抑制的敏感性。用 Src 抑制剂萨拉卡替尼 (AZD0530) 处理四种“Met 成瘾”胃癌细胞系（GTL16、MKN45、HS746T 和 SNU5）和三种不依赖 Met 的胃癌细胞系（KATO III、AGS 和 NCI-N87）。在GTL16和KATO III中，Src中和也是通过达沙替尼和RNA干扰实现的。使用抗磷蛋白抗体和寡核苷酸微阵列探讨了 Src 抑制的生化和转录后果。 结果：在 Met 成瘾的胃癌细胞系中抑制 Src (a) 降低了参与细胞增殖和细胞凋亡保护的信号中间体的磷酸化/激活水平，并下调了几种细胞周期调节因子的表达； (b) 减少不依赖锚定的生长； (c) Met抑制导致的细胞活力受损增强； (d) 异种移植模型中延迟肿瘤发生。萨拉卡替尼全身治疗后肿瘤异种移植组织的免疫组织化学分析显示，肿瘤细胞增殖指数降低，细胞凋亡增加，磷酸焦点粘附激酶和磷酸桩蛋白染色减少。肿瘤基质参数如血管生成或炎症浸润不受影响。在克隆形成存活测定中，未对Met成瘾的胃癌细胞对Src抑制的敏感性低于对Met成瘾的细胞。结论：Src是Met驱动的肿瘤生长的关键下游转导器。靶向 Src 可能为 Met 成瘾肿瘤提供治疗益处。临床癌症研究； 16（15）； 3933-43。 (C) 2010 AACR。
Purpose: We examined whether inhibition of Src tyrosine kinase, a downstream effector of the MET oncogene, can hinder the malignant properties of gastric tumors dependent on Met for growth and survival.Experimental Design: Sensitivity to Src inhibition was determined in vitro by measuring clonogenic survival (anchorage-independent growth) and in vivo by establishing xenograft models. Four "Met-addicted" gastric carcinoma cell lines (GTL16, MKN45, HS746T, and SNU5) and three Met-independent gastric carcinoma cell lines (KATO III, AGS, and NCI-N87) were treated with the Src inhibitor saracatinib (AZD0530). In GTL16 and KATO III, Src neutralization was also achieved by dasatinib and RNA interference. The biochemical and transcriptional consequences of Src inhibition were explored using anti-phosphoprotein antibodies and oligonucleotide microarrays.Results: Inhibition of Src in Met-addicted gastric carcinoma cell lines (a) decreased the phosphorylation/activation levels of signaling intermediates involved in cell proliferation and protection from apoptosis and down-modulated the expression of several cell cycle regulators; (b) reduced anchorage-independent growth; (c) enhanced impairment of cell viability produced by Met inhibition; and (d) delayed tumorigenesis in xenotransplantation models. Immunohistochemical analysis of tumor xenograft tissues following systemic treatment with saracatinib showed a reduction of tumor cell proliferation index, increased apoptosis, and diminished phospho-focal adhesion kinase and phospho-paxillin staining. Tumor stroma parameters such as angiogenesis or inflammatory infiltration were unaffected. In clonogenic survival assays, gastric carcinoma cells without addiction to Met were less sensitive than Met-addicted cells to Src inhibition.Conclusions: Src is as a key downstream transducer of Met-driven tumor growth. Targeting Src might provide therapeutic benefit in Met-addicted tumors. Clin Cancer Res; 16(15); 3933-43. (C) 2010 AACR.