CAJ | 학술논문

目的：针对非小细胞肺癌（NSCLC）患者组织标本鼠类肉瘤病毒癌基因同源基因（KRAS）和鼠类肉瘤滤过性毒菌致癌同源体 B1（BRAF）驱动基因突变使用特异引物双扩增实时蝎形探针扩增阻滞突变系统（sARMS-PCR）检测的可行性；了解 KRAS 和 BRAF 基因突变患者临床病理特征，为NSCLC 患者个体化治疗提供理论依据。方法收集89例NSCLC 患者肿瘤组织甲醛固定石蜡包埋标本（FFPE），采用FFPE 样品 DNA 分离试剂盒（离心柱型）提取 DNA，使用sARMS-PCR 同时进行 KRAS 及 BRAF 基因突变检测。结果① KRAS 基因突变21例（21／89）；其中KRAS 基因7种热点突变中，检出6种热点突变，均位于第12、13位密码子，1例同时检出存在G12D 及G12V 位点突变，1例同时检出存在G12C 及G12V 位点突变；未发现G12S 突变型；② BRAF 基因突变1例（1／89），突变位点为V600E，为女性，黏液腺癌；③未见KRAS 和BRAF 基因同时突变现象。结论临床使用sARMS-PCR 技术检测NSCLC 患者KRAS 和BRAF 基因突变有较强敏感性，且石蜡组织标本取材方便，可以作为两种基因的临床检测方法；KRAS 和BRAF 基因突变与年龄、吸烟史、病理分型等均无明显相关性，KRAS 基因突变与性别相关，男性高于女性；KRAS 与BRAF 基因是独立存在的。
목적：침대비소세포폐암（NSCLC）환자조직표본서류육류병독암기인동원기인（KRAS）화서류육류려과성독균치암동원체 B1（BRAF）구동기인돌변사용특이인물쌍확증실시갈형탐침확증조체돌변계통（sARMS-PCR）검측적가행성；료해 KRAS 화 BRAF 기인돌변환자림상병리특정，위NSCLC 환자개체화치료제공이론의거。방법수집89례NSCLC 환자종류조직갑철고정석사포매표본（FFPE），채용FFPE 양품 DNA 분리시제합（리심주형）제취 DNA，사용sARMS-PCR 동시진행 KRAS 급 BRAF 기인돌변검측。결과① KRAS 기인돌변21례（21／89）；기중KRAS 기인7충열점돌변중，검출6충열점돌변，균위우제12、13위밀마자，1례동시검출존재G12D 급G12V 위점돌변，1례동시검출존재G12C 급G12V 위점돌변；미발현G12S 돌변형；② BRAF 기인돌변1례（1／89），돌변위점위V600E，위녀성，점액선암；③미견KRAS 화BRAF 기인동시돌변현상。결론림상사용sARMS-PCR 기술검측NSCLC 환자KRAS 화BRAF 기인돌변유교강민감성，차석사조직표본취재방편，가이작위량충기인적림상검측방법；KRAS 화BRAF 기인돌변여년령、흡연사、병리분형등균무명현상관성，KRAS 기인돌변여성별상관，남성고우녀성；KRAS 여BRAF 기인시독립존재적。
Objective To explore the feasibility of use scorpion probe amplification refractory mutation system (sARMS-PCR)to detect Kirsten rat sarcoma viral oncogene homolog(KRAS)and V-rafmurine sarcoma viral onco-gene homologB1(BRAF)gene mutations in non-small cell lung cancer(NSCLC)patients.To realize the clinical and pathological feature of KRAS,BRAF gene mutations in NSCLC patients,and to provide a theoretical basis for individualized treatment of NSCLC patients.Methods We collected 89 cases of tumor formalin-fixed paraffin-em-bedded tissue specimens (FFPE)in NSCLC patients,then used DNA isolation kit to extract DNA,and used sARMS to test KRAS and BRAF gene mutation.Results ① We found KRAS mutations in 21 patients(21 /89),the muta-tion rate was 23.6%;the KRAS gene mutations included seven kinds of hot spots mutations,in which 6 were loca-ted at codon 12 and 13;the G12D and G12V site mutation and the G12C and G12C were detected from the 6 hot spots respectively;G12S mutant was not found;② We only found BRAF gene mutation in one patient(1 /89),the mutation rate was 1.1%.The mutation site was located at V600E,was a women,mucinous adenocarcinoma pa-tient;③ KRAS and BRAF gene never co-existed in the same patient.Conclusion Using sARMS-PCR technology to detect KRAS and BRAF gene mutations is feasible in NSCLC patients.There is no significant correlation with age,smoking history,pathological type in KRAS and BRAF gene mutations in NSCLC patients.KRAS gene muta-tion is related to gender,more common in women than in men(P <0.05);KRAS and BRAF gene never coexist in the same patient.