Peptide nucleic acid-locked nucleic acid polymerase chain reaction clamp-based detection test for gefitinib-refractory T790M epidermal growth factor receptor mutation
Peptide nucleic acid-locked nucleic acid polymerase chain reaction clamp-based detection test for gefitinib-refractory T790M epidermal growth factor receptor mutation
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DOI：
10.1111/j.1349-7006.2007.00706.x
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发表时间：
2008-03-01
期刊：
影响因子：
5.7
通讯作者：
Hagiwara, Koichi
中科院分区：
文献类型：
--
作者：
Miyazawa, Hitoshi;Tanaka, Tomoaki;Hagiwara, Koichi
在部分非小细胞肺癌(NSCLS)中观察到表皮生长因子受体(EGFR)的突变。EGFR突变阳性的非小细胞肺癌对吉非替尼有反应。继发性T790M突变使吉非替尼对非小细胞肺癌耐药。设计了一种基于肽核酸锁定核酸聚合酶链式反应钳夹法的T790M突变检测方法。试验的特异度和敏感度均大于0.99。检测显示，只有一小部分PC-13细胞携带T790M突变。检测还显示，在吉非替尼治疗前获得的151例非小细胞肺癌标本中，没有一例发现T790M突变，而在从非小细胞肺癌获得的4例对吉非替尼无效的标本中，有4例发现了T790M突变。在一例L858R阳性的EGFR等位基因被扩增为多个副本的患者中，在吉非替尼治疗期间出现了L858R-T790M双突变等位基因。该等位基因高表达。基于多肽核酸锁定核酸聚合酶链式反应钳夹法的T790M突变检测方法灵敏、特异，适用于临床实践。它在吉非替尼治疗过程中检测到T790M阳性细胞，从而有助于设计有效的治疗T790M阳性非小细胞肺癌的方法。
Mutations in the epidermal growth factor receptor (EGFR) are observed in a fraction of non-small-cell lung cancers (NSCLS). EGFR mutation-positive NSCLS responds to gefitinib. Secondary T790M mutation confers gefitinib resistance to NSCLS. A detection test for the T790M mutation was designed based on the peptide nucleic acid-locked nucleic acid polymerase chain reaction clamp method. The specificity and sensitivity of the test were both greater than 0.99. The test revealed that only a small population of the PC-13 cells carried the T790M mutation. The test also revealed that the T790M mutation was found in none of 151 NSCLC specimens obtained before gefitinib treatment, whereas it was found in four of four specimens obtained from NSCLS that had become refractory to gefitinib. In one patient in whom the L858R-positive EGFR allele was amplified to multiple copies, an L858R-T790M double-mutant allele emerged during the gefitinib therapy. This allele was expressed highly. The T790M mutation detection test based on the peptide nucleic acid-locked nucleic acid polymerase chain reaction clamp method is sensitive and specific, and is applicable to clinical practice. It detects T790M-positive cells in the course of gefitinib treatment, and thus will help to devise therapies effective for T790M-positive NSCLS.