Functional expression and mutations of c-met and its therapeutic inhibition with SU11274 and small interfering RNA in non-small cell lung cancer
Functional expression and mutations of c-met and its therapeutic inhibition with SU11274 and small interfering RNA in non-small cell lung cancer
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DOI：
10.1158/0008-5472.can-04-2650
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发表时间：
2005-02-15
期刊：
影响因子：
11.2
通讯作者：
Salgia, R
中科院分区：
文献类型：
--
作者：
Ma, PC;Jagadeeswaran, R;Salgia, R
非小细胞肺癌(NSCLC)是一种难治性疾病。C-Met受体是一种有吸引力的潜在靶点，可用于治疗人类癌症。我们提供了强有力的证据表明，c-Met在非小细胞肺癌细胞系和肿瘤组织中过表达、激活，有时还发生突变。C-Met在所有(100%)NSCLC肿瘤组织(n=23)和大部分(89%)细胞系(n=9)中均有表达。61%的肿瘤组织总c-Met呈强阳性表达，尤其是腺癌(67%)。免疫组织化学检测到磷酸化蛋氨酸(p-Met)[Y1003]和[Y1230/1234/1235]的特异性表达。P-Met主要在NSCLC肿瘤侵袭前沿表达。在NSCLC细胞系和腺癌组织的信号素结构域(E168D、L299F、S323G和N375S)和膜旁结构域(R988C、R988C+T1010I、S1058P和跳过整个膜旁结构域的选择性剪接产物)中发现了C-Met改变。我们通过小干扰RNA下调非小细胞肺癌细胞中受体表达50%至60%，证实c-Met是潜在的治疗靶点。这导致p-Met和磷酸化AKT受到抑制，72小时后细胞存活率抑制达57.1+/-7.2%。选择性小分子抑制剂c-Met SU11274抑制表达c-Met的非小细胞肺癌细胞的活力。SU11274还可阻断肝细胞生长因子诱导的c-Met的磷酸化及其下游信号转导。在这里，我们首次通过小干扰RNA靶向和小分子抑制剂提供了c-Met在非小细胞肺癌生物学和生化中的重要作用的直接证据。这些结果表明，抑制c-Met将是治疗NSCLC的重要策略，以改善其临床预后。
Non-small cell lung cancer (NSCLC) is a difficult disease to treat. The c-Met receptor is an attractive potential target for novel therapeutic inhibition in human cancers. We provide strong evidence that c-Met is overexpressed, activated, and sometimes mutated in NSCLC cell lines and tumor tissues. Expression of c-Met was found in all (100%) of the NSCLC tumor tissues examined (n = 23) and most (89%) of the cell lines (n = 9). Sixty-one percent of tumor tissues strongly expressed total c-Met, especially adenocarcinoma (67%). Specific expression of phospho-Met (p-Met) [Y1003] and [Y1230/1234/1235] was seen by immunohistochemistry. p-Met expression was preferentially observed at the NSCLC tumor invasive fronts. c-Met alterations were identified within the semaphorin domain (E168D, L299F, S323G, and N375S) and the juxtamembrane domain (R988C, R988C + T1010I, S1058P, and alternative splice product skipping entire juxtamembrane domain) of a NSCLC cell line and adenocarcinoma tissues. We validated c-Met as potential therapeutic target using small interfering RNA down-regulation of the receptor expression by 50% to 60% in NSCLC cells. This led to inhibition of p-Met and phospho-AKT and up to 57.1 +/- 7.2% cell viability inhibition at 72 hours. The selective small molecule inhibitor of c-Met SU11274 inhibited cell viability in c-Met-expressing NSCLC cells. SU11274 also abrogated hepatocyte growth factor-induced phosphorylation of c-Met and its downstream signaling. Here, we pro-Ode first direct evidence by small interfering RNA targeting and small molecule inhibitor that c-Met is important in NSCLC biology and biochemistry. These results indicate that c-Met inhibition will be an important therapeutic strategy against NSCLC to improve its clinical outcome.