The mutation rates of EGFR in non-small cell lung cancer and KRAS in colorectal cancer of Chinese patients as detected by pyrosequencing using a novel dispensation order.
The mutation rates of EGFR in non-small cell lung cancer and KRAS in colorectal cancer of Chinese patients as detected by pyrosequencing using a novel dispensation order.
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使用新型配药顺序焦磷酸测序检测中国患者非小细胞肺癌EGFR和结直肠癌KRAS突变率
DOI：
10.1186/s13046-015-0179-9
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发表时间：
2015-06-18
期刊：
影响因子：
--
通讯作者：
Shen L
中科院分区：
文献类型：
--
作者：
Xie G;Xie F;Wu P;Yuan X;Ma Y;Xu Y;Li L;Xu L;Yang M;Shen L
本研究的目的是建立一种经济有效的方法来检测福尔马林固定石蜡包埋(FFPE)的非小细胞肺癌(NSCLC)和结直肠癌(CRC)标本中的EGFR和KRAS突变。另外，对494例非小细胞肺癌和1099例结直肠癌患者标本进行PS检测，根据临床病理特征评价EGFR或KRAS突变模式。35.63%的非小细胞肺癌标本中检测到EGFR突变，39.76%的结直肠癌标本中检测到KRAS突变。经多因素分析发现，65岁及以下(OR = 2.51)、有非吸烟史(OR = 3.63)、腺癌(OR = 3.57)的非小细胞肺癌患者EGFR基因突变显著，而女性患者无显著差异(OR = 0.64)。女性患者(OR = 1.64)、50岁以上患者(OR = 4.17)、腺癌患者(OR = 2.41)中KRAS基因突变的发生率较高。结论首次通过检测EGFR18-21外显子突变和KRAS基因外显子2突变，对结直肠癌标本中PS进行了广泛验证。我们的结果证明了PS分析在检测临床样本中体细胞EGFR和KRAS突变方面的应用，并为中国人NSCLC和CRC提供了重要的临床和分子特征。
BackgroundThe purpose of this study was to develop a cost-effective approach for the determination of EGFR and KRAS mutations in formalin-fixed paraffin-embedded (FFPE) non-small cell lung cancer (NSCLC) and colorectal cancer (CRC) samples from Chinese patients based on a sensitive pyrosequencing (PS) technique.MethodsThe NSCLC and CRC cell lines were tested to determine the limitation of detection and reproducibility of the PS method. In addition, 494 NSCLC and 1099 CRC patient samples were assayed by PS to evaluate the EGFR or KRAS mutation patterns according to the clinicopathological features.ResultsThe PS assay was able to reproducibly detect as few as 2 % mutant alleles with excellent linearity. EGFR mutations were detected in 35.63 % of the NSCLC samples, and KRAS mutations were detected in 39.76 % of the CRC samples. EGFR mutations were more frequently observed to be significant by multivariate analysis in NSCLC patients who were 65 years old or younger (OR = 2.51), had a nonsmoking history (OR = 3.63), and adenocarcinoma (OR = 3.57), but not in females (OR = 0.64). KRAS mutations were more frequently detected in CRC patients who were female (OR = 1.64) and 50 years old or older (OR = 4.17), and had adenocarcinoma (OR = 2.41).ConclusionsThis is the first extensive validation of PS on FFPE samples using the detection of EGFR exons 18–21 mutations and KRAS exon 2 mutations. Our results demonstrate the utility of PS analysis for the detection of somatic EGFR and KRAS mutations in clinical samples and provide important clinical and molecular characteristics of NSCLC and CRC from Chinese patients.