Clinical significance of multiple gene detection with a 22-gene panel in formalin-fixed paraffin-embedded specimens of 207 colorectal cancer patients
Clinical significance of multiple gene detection with a 22-gene panel in formalin-fixed paraffin-embedded specimens of 207 colorectal cancer patients
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22基因组检测207例结直肠癌福尔马林固定石蜡包埋标本中多基因的临床意义
DOI：
10.1007/s10147-018-1377-1
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发表时间：
2019-02-01
影响因子：
3.3
通讯作者：
Zhang, Wei
中科院分区：
文献类型：
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作者：
Gao, Xian Hua;Yu, Guan Yu;Zhang, Wei
背景同时检测多个分子生物标志物有助于预测结直肠癌（CRC）患者的治疗反应和预后。方法采用基于下一代测序（NGS）的多重PCR技术，对207例CRC患者福尔马林固定石蜡包埋（FFPE）组织标本中的103个热点区域组成的22个基因组进行检测。这22个基因包括AKT 1、ALK、BRAF、CTNNB 1、DDR2、EGFR、ERBB 2、ERBB 4、FBXW 7、FGFR 1、FGFR 2、FGFR 3、KRAS、MAP 2K 1、MET、NOTCH 1、NRAS、PIK 3CA、PTEN、SMAD 4、STK 11和TP 53。在本研究中鉴定的总共414个变体中，384个、25个和5个是单核苷酸变体、缺失和插入。前四个频繁突变的基因是TP 53、KRAS、PIK 3CA和FBXW 7。NGS-PCR技术检测KRAS和BRAF突变的结果与常规ARMS-PCR检测结果高度一致。单因素和多因素分析显示，TNM分期、血清CEA升高、总变异数≥ 2、AKT 1和PTEN突变是DFS缩短的独立预测因子;分化差、TNM分期、总变异数≥ 2、BRAF、CTNNB 1和NRAS突变是OS缩短的独立预测因子。TNM分期和总变异数≥ 2是预测DFS和OS的独立指标，多基因突变检测可为结直肠癌患者的TNM分期提供额外的预后信息。
BackgroundSimultaneous detection of multiple molecular biomarkers is helpful in the prediction of treatment response and prognosis for colorectal cancer (CRC) patients.MethodsA 22-gene panel consisting of 103 hotspot regions was utilized in the formalin-fixed paraffin-embedded (FFPE) tissue samples of 207 CRC patients, using the next-generation sequencing (NGS)-based multiplex PCR technique. Those 22 genes included AKT1, ALK, BRAF, CTNNB1, DDR2, EGFR, ERBB2, ERBB4, FBXW7, FGFR1, FGFR2, FGFR3, KRAS, MAP2K1, MET, NOTCH1, NRAS, PIK3CA, PTEN, SMAD4, STK11, and TP53.ResultsOf the 207 patients, 193 had one or more variants, with 170, 20, and 3 having one, two, and three mutated genes, respectively. Of the total 414 variants identified in this study, 384, 25, and 5 were single-nucleotide variants, deletion, and insertion. The top four frequently mutated genes were TP53, KRAS, PIK3CA, and FBXW7. There was high consistency between the results of NGS–PCR technique and routine ARMS-PCR in KRAS and BRAF mutation detection. Univariate and multivariate analyses demonstrated that advanced TNM stage, elevated serum CEA, total variants number ≥ 2, AKT1 and PTEN mutation were independent predictors of shorter DFS; poor differentiation, advanced TNM stage, total variants number ≥ 2, BRAF, CTNNB1 and NRAS mutation were independent predictors of shorter OS.ConclusionsIt is feasible to detect multiple gene mutations with a 22-gene panel in FFPE CRC specimens. TNM stage and total variants number ≥ 2 were independent predictors of DFS and OS. Detection of multiple gene mutations may provide additional prognostic information to TNM stage in CRC patients.