The use of a two-tiered testing strategy for the simultaneous detection of small EGFR mutations and EGFR amplification in lung cancer.
The use of a two-tiered testing strategy for the simultaneous detection of small EGFR mutations and EGFR amplification in lung cancer.
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DOI：
10.1371/journal.pone.0117983
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Kozlowski P
中科院分区：
文献类型：
--
作者：
Lewandowska MA;Czubak K;Klonowska K;Jozwicki W;Kowalewski J;Kozlowski P
肺癌是全球癌症相关死亡的主要原因。由于对疾病的分子机制有了更好的了解，以及对生物标志物的识别使得更具体的癌症治疗成为可能，肺癌的诊断和治疗取得了最近的进展。个体化治疗最著名的例子之一是使用酪氨酸激酶抑制剂，如吉非替尼和厄洛替尼，成功治疗基于特定EGFR突变选择的非小细胞肺癌患者。因此，突变的可靠检测对于适当治疗的应用至关重要。在这项研究中，我们测试了一种双层突变检测策略，使用实时PCR检测作为一种经过验证的高灵敏度方法，以及基于多重连接依赖探针扩增（MLPA）的EGFRmut+检测作为二级标准灵敏度方法。应用MLPA方法的另一个优点是，它允许同时检测EGFR突变和EGFR、MET和ERBB2中的拷贝数改变（即扩增）。我们的分析表明，这两种方法之间具有高度的一致性。通过使用这种双层策略，我们可靠地确定了200多个肺癌样本中EGFR突变和EGFR、MET和ERBB2扩增的频率。此外，利用同时进行的拷贝数和小突变分析，我们发现EGFR突变与EGFR扩增之间存在很强的相关性，并且EGFR突变/扩增与MET和ERBB2扩增具有互斥性。我们的结果证明了两层EGFR检测策略的可靠性和有效性。
Lung cancer is the leading cause of cancer-related death worldwide. Recent progress in lung cancer diagnosis and treatment has been achieved due to a better understanding the molecular mechanisms of the disease and the identification of biomarkers that allow more specific cancer treatments. One of the best known examples of personalized therapy is the use of tyrosine kinase inhibitors, such as gefitinib and erlotinib, for the successful treatment of non-small-cell lung cancer patients selected based on the specific EGFR mutations. Therefore, the reliable detection of mutations is critical for the application of appropriate therapy. In this study, we tested a two-tiered mutation detection strategy using real-time PCR assays as a well-validated high-sensitivity method and multiplex ligation-dependent probe amplification (MLPA)-based EGFRmut+ assay as a second-tier standard-sensitivity method. One additional advantage of the applied MLPA method is that it allows the simultaneous detection of EGFR mutations and copy-number alterations (i.e., amplifications) in EGFR, MET and ERBB2. Our analysis showed high concordance between these two methods. With the use of this two-tier strategy, we reliably determined the frequency of EGFR mutations and EGFR, MET and ERBB2 amplifications in over 200 lung cancer samples. Additionally, taking advantage of simultaneous copy number and small mutation analyses, we showed a very strong correlation between EGFR mutations and EGFR amplifications and a mutual exclusiveness of EGFR mutations/amplifications with MET and ERBB2 amplifications. Our results proved the reliability and usefulness of the two-tiered EGFR testing strategy.
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影响因子：
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作者：
Marcinkowska, Malgorzata;Wong, Kwok-Kin;Kozlowski, Piotr
通讯作者：
Kozlowski, Piotr
影响因子：
56.9
作者：
Gonzalez, E;Kulkarni, H;Ahuja, SK
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Ahuja, SK
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158.5
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Kobayashi, S;Boggon, TJ;Halmos, B
通讯作者：
Halmos, B
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作者：
Hwang, Vicki J.;Maar, Dianna;Tassone, Flora
通讯作者：
Tassone, Flora
影响因子：
8
作者：
Gazdar, A. F.
通讯作者：
Gazdar, A. F.