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首页> 外文期刊>International Journal of Molecular Sciences >KRAS Mutation Detection in Paired Frozen and Formalin-Fixed Paraffin-Embedded (FFPE) Colorectal Cancer Tissues
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KRAS Mutation Detection in Paired Frozen and Formalin-Fixed Paraffin-Embedded (FFPE) Colorectal Cancer Tissues

机译：配对的冷冻和福尔马林固定石蜡包埋（FFPE）大肠癌组织中的KRAS突变检测

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摘要

KRAS mutation has been unambiguously identified as a marker of resistance to cetuximab-based treatment in metastatic colorectal cancer (mCRC) patients. However, most studies of KRAS mutation analysis have been performed using homogenously archived CRC specimens, and studies that compare freshly frozen specimens and formalin-fixed paraffin-embedded (FFPE) specimens of CRC are lacking. The aim of the present study was to evaluate the impact of tissue preservation on the determination of KRAS mutational status. A series of 131 mCRC fresh-frozen tissues were first analyzed using both high-resolution melting (HRM) and direct sequencing. KRAS mutations were found in 47/131 (35.8%) using both approaches. Out of the 47 samples that were positive for KRAS mutations, 33 had available matched FFPE specimens. Using HRM, 2/33 (6%) demonstrated suboptimal template amplification, and 2/33 (6%) expressed an erroneous wild-type KRAS profile. Using direct sequencing, 6/33 (18.1%) displayed a wild-type KRAS status, and 3/33 (9.1%) showed discordant mutations. Finally, the detection of KRAS mutations was lower among the FFPE samples compared with the freshly frozen samples, demonstrating that tissue processing clearly impacts the accuracy of KRAS genotyping.
机译：在转移性结直肠癌（mCRC）患者中，已明确确定KRAS突变是对西妥昔单抗治疗耐药的标志物。但是，大多数KRAS突变分析的研究都是使用均一存档的CRC标本进行的，缺乏将新鲜冷冻的标本和福尔马林固定石蜡包埋的（FFPE）标本进行比较的研究尚缺乏。本研究的目的是评估组织保存对KRAS突变状态测定的影响。首先使用高分辨率熔解（HRM）和直接测序技术对一系列131 mCRC新鲜冷冻组织进行了分析。两种方法均在47/131（35.8％）中发现KRAS突变。在KRAS突变阳性的47个样本中，有33个具有匹配的FFPE标本。使用HRM，2/33（6％）表现出次佳的模板扩增，而2/33（6％）表达了错误的野生型KRAS谱。使用直接测序，6/33（18.1％）表现出野生型KRAS状态，3/33（9.1％）表现出不一致的突变。最后，与新鲜冷冻的样品相比，FFPE样品中的KRAS突变检测率更低，这表明组织加工明显影响了KRAS基因分型的准确性。

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