Detection of KRAS Mutations in Plasma DNA Using a fully Automated Rapid Detection System in Colorectal Cancer Patients
Detection of KRAS Mutations in Plasma DNA Using a fully Automated Rapid Detection System in Colorectal Cancer Patients
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DOI：
10.1007/s12253-016-0175-1
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发表时间：
2017-01
影响因子：
2.8
通讯作者：
Kazuhisa Hosoya;S. Matsusaka;T. Kashiwada;Koichi Suzuki;N. Ureshino;A. Sato;Y. Miki;Kazuki Kitera;M. Hirai;K. Hatake;S. Kimura;N. Sueoka-Aragane
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文献类型：
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作者：
Kazuhisa Hosoya;S. Matsusaka;T. Kashiwada;Koichi Suzuki;N. Ureshino;A. Sato;Y. Miki;Kazuki Kitera;M. Hirai;K. Hatake;S. Kimura;N. Sueoka-Aragane
KRAS突变已被认为是结直肠癌患者对抗EGFR抗体的原发性耐药的预测标志物。此外，新近发现的KRAS突变与抗EGFR抗体化疗获得性耐药有关。考虑到这一证据，KRAS突变的监测对于做出治疗决定是必不可少的，并且该方法应该是无创的，允许重复检查。最近，我们建立了一种新的KRAS突变的自动化检测系统，称为突变偏置PCR淬灭探针系统（MBP-QP）。我们研究的目标是调查抗EGFR抗体治疗期间监测KRAS突变的潜力。用含有KRAS突变的对照质粒检测MBP-QP的最低检出量为1-9个拷贝，在野生型和突变型混合质粒中检出率为0.05-0.3%。采用MBP-QP和聚合酶链反应反向序列特异性寡核苷酸（PCR-rSSO）对120例结直肠癌患者进行KRAS突变基因分型。MBP-QP和PCR-rSSO检测血浆DNA与癌组织的符合率分别为68%和66%，说明这两种检测系统在检测血浆DNA中KRAS突变方面是相当的。需要进行前瞻性研究以明确血浆DNA中检测到的KRAS突变是否是抗EGFR抗体治疗疗效的预测标志物。
KRASmutations have been recognized as predictive markers of primary resistance to anti-EGFR-antibodies in colorectal cancer patients. In addition, newly detectedKRASmutations have been reported to be related with acquired resistance to chemotherapy containing anti-EGFR antibody. Considering this evidence, monitoring ofKRASmutations is indispensable for making treatment decisions, and the method should be non-invasive allowing repeated examinations. Recently, we established a novel automated sensitive detection system forKRASmutations, named mutation-biased PCR quenching probe system (MBP-QP). The goal of our study was to investigate the potential for monitoring KRAS mutations during treatment with anti-EGFR antibodies. The detection limit of MBP-QP using a control plasmid containingKRASmutations was 1–9 copies, and 0.05–0.3% mutant plasmid was detectable in a mixture of wild type and mutants. One-hundred twenty colorectal cancer patients were genotyped forKRASmutations with MBP-QP as well as polymerase chain reaction reverse sequence-specific oligonucleotide (PCR-rSSO), which has already been applied to cancer tissue samples in the clinical setting. Concordance rates between plasma DNA and cancer tissues were 68% with MBP-QP and 66% with PCR-rSSO, indicating that these systems are equivalent in terms of detectingKRASmutations with plasma DNA.KRASmutations in plasma DNA were frequently observed in systemic metastatic cancer patients, and in three patientsKRASmutations appeared after chemotherapy containing anti-EGFR antibody. A prospective study is needed for clarifying whetherKRASmutations detected in plasma DNA are predictive markers of treatment efficacy with anti-EGFR antibody.