MET gene amplification or EGFR mutation activate MET in lung cancers untreated with EGFR tyrosine kinase inhibitors.
MET gene amplification or EGFR mutation activate MET in lung cancers untreated with EGFR tyrosine kinase inhibitors.
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DOI：
10.1002/ijc.24150
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发表时间：
2009-04-15
影响因子：
6.4
通讯作者：
Toyooka, Shinichi
中科院分区：
文献类型：
--
作者：
Kubo, Takafumi;Yamamoto, Hiromasa;Lockwood, William W.;Valencia, Ilse;Sob, Junichi;Peyton, Michael;Jida, Masaru;Otani, Hiroki;Fujii, Tetsuva;Ouchida, Mamoru;Takigawa, Nagio;Kiura, Katsuyuki;Shimizu, Kenji;Date, Hiroshi;Minna, John D.;Varella-Garcia, Marileila;Lam, Wan L.;Gazdar, Adi F.;Toyooka, Shinichi
我们分析了未经EGFR酪氨酸激酶抑制剂(TKIs)治疗的伴有或不伴有EGFR突变的NSCLC中MET蛋白和拷贝数。应用实时定量聚合酶链式反应技术检测了28例非小细胞肺癌、4株人支气管上皮细胞系(HBEC)和100例原发肿瘤中MET基因的拷贝数。芯片比较基因组杂交和荧光原位杂交证实阳性结果。用Western印迹法检测24株NSCLC和2株HBEC细胞中总蛋白和磷酸化MET蛋白的表达。检测EGFR基因外显子19缺失、T790M和L858R突变。用siRNA敲除EGFR，以检测EGFR与MET活化的关系。在28个NSCLC细胞系中有3个观察到高水平的MET扩增，在100个未经EGFR-TKI治疗的原发肺肿瘤中有2个观察到高水平的MET扩增。MET蛋白在3株MET高扩增的细胞系中均有高表达和磷酸化。而在21个未扩增MET的非小细胞肺癌细胞系中，有6个细胞系显示有磷酸化MET(p=0.042)。此外，6株表达磷酸化MET但未扩增MET的细胞系均为EGFR突变株(p=0.0039)。在3个未扩增出MET基因拷贝数的EGFR突变细胞系中，siRNA介导的EGFR基因敲除使其磷酸化MET表达消失。相反，2株MET基因扩增的细胞株的磷酸化MET表达不受EGFR基因敲除的影响。结果表明，未经处理的非小细胞肺癌中存在MET扩增，而非小细胞肺癌中存在EGFR突变或MET扩增激活的MET蛋白。
We analyzed MET protein and copy number in NSCLC with or without EGFR mutations untreated with EGFR tyrosine kinase inhibitors (TKIs). MET copy number was examined in 28 NSCLC and 4 human bronchial epithelial cell lines (HBEC) and 100 primary tumors using quantitative real-time PCR. Positive results were confirmed by array comparative genomic hybridization and fluorescence in-situ hybridization. Total and phospho-MET protein expression was determined in 24 NSCLC and 2 HBEC cell lines using Western blot. EGFR mutations were examined for exon 19 deletions, T790M, and L858R. Knockdown of EGFR with siRNA was performed to examine the relation between EGFR and MET activation. High-level MET amplification was observed in 3 of 28 NSCLC cell lines and in 2 of 100 primary lung tumors that had not been treated with EGFR-TKIs. MET protein was highly expressed and phosphorylated in all the 3 cell lines with high MET amplification. In contrast, 6 NSCLC cell lines showed phospho-MET among 21 NSCLC cell lines without MET amplification (p = 0.042). Furthermore, those 6 cell lines harboring phospho-MET expression without MET amplification were all EGFR mutant (p = 0.0039). siRNA-mediated knockdown of EGFR abolished phospho-MET expression in examined 3 EGFR mutant cell lines of which MET gene copy number was not amplified. By contrast, phospho-MET expression in 2 cell lines with amplified MET gene was not down-regulated by knockdown of EGFR. Our results indicated that MET amplification was present in untreated NSCLC and EGFR mutation or MET amplification activated MET protein in NSCLC.
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影响因子：
158.5
作者：
Lynch, TJ;Bell, DW;Haber, DA
通讯作者：
Haber, DA
影响因子：
6.4
作者：
Lockwood, William W.;Coe, Bradley P.;Lam, Wan L.
通讯作者：
Lam, Wan L.
影响因子：
64.8
作者：
GIORDANO, S;PONZETTO, C;COMOGLIO, PM
通讯作者：
COMOGLIO, PM
DOI：
10.1073/pnas.0508776103
发表时间：
2006-02-14
影响因子：
11.1
作者：
Smolen, GA;Sordella, R;Haber, DA
通讯作者：
Haber, DA
DOI：
10.1073/pnas.0502860102
发表时间：
2005-05-24
影响因子：
11.1
作者：
Kwak, EL;Sordella, R;Haber, DA
通讯作者：
Haber, DA