肺腺癌细胞学标本十基因联合检测分析  

Ten-gene panel detection for cytological specimens of lung adenocarcinoma

作　　者：张雪莲 程凯 饶秋 ZHANG Xuelian;CHENG Kai;RAO Qiu(Jinling Clinical Medical College of Nanjing Medical University,Nanjing 210002,China;Nanjing KingMed Diagnostics Center,Nanjing 210032,China)

机构地区：[1]南京医科大学金陵临床医学院,南京210002 [2]南京金域医学检验所有限公司,南京210032

出　　处：《临床与实验病理学杂志》2024年第10期1033-1040,共8页Chinese Journal of Clinical and Experimental Pathology

摘　　要：目的探讨分子检测在肺腺癌细胞蜡块标本中的应用及临床意义。方法收集200例肺腺癌细胞蜡块标本,其中49例行十基因联合检测(包括EGFR、EML4-ALK、ROS1、KRAS、HER2、BRAF、RET、MET、NRAS和PIK3CA),151例行EGFR单基因检测;97例行Ventana免疫组化染色检测EML4-ALK融合基因突变情况,并与PCR检测结果进行对比。总结突变特点并观察突变和融合病例的细胞形态学特征。结果49例十基因联合检测驱动基因总突变率为65.31%,与99例肺腺癌组织标本十基因联合检测驱动基因总突变率(79.8%)相比差异无统计学意义(P>0.05),其中ALK、ROS1和RET融合突变率分别为4.08%、4.08%和2.04%,EGFR、KRAS、HER2、MET和BRAF突变率分别为38.78%、6.12%、4.08%、4.08%和2.04%。151例EGFR单基因突变率为54.97%,其中男性患者的EGFR单基因突变率为42.86%,女性患者的EGFR单基因突变率为67.57%,差异有统计学意义(P<0.05);EGFR基因的19号外显子缺失突变(19-Del)和21号外显子点突变(21-L858R)占比最高,分别为54.22%和37.35%,显著高于其他突变类型(P<0.001)。免疫组化检测97例细胞蜡块EML4-ALK蛋白阳性率为15.46%,其中6例弱阳性(+)的分子检测结果均为阴性,9例弥漫阳性()的分子检测结果均为阳性,免疫组化弥漫强阳性者与分子检测结果高度一致(Kappa=1)。观察基因改变阳性病例细胞学形态特征,发现KARS突变者特征较为特殊,其肿瘤细胞散在或呈小团状排列,胞质空泡更多见。结论可用肺腺癌细胞学标本进行分子检测,其敏感性较高,多基因联合检测优于单基因检测;EML4-ALK融合基因可采用免疫组化法进行筛选;KRAS突变的肺腺癌具有特殊的细胞学形态。Purpose To explore the application and clinical significance of molecular detection in cell block specimens of lung adenocarcinoma.Methods A total of 200 cell block specimens of lung adenocarcinoma were collected.Among them 49 specimens underwent multi-gene panel detection(including EGFR,EML4-ALK,ROS1,KRAS,HER2,BRAF,RET,MET,NRAS,and PIK3CA),while 151 specimens underwent EGFR single-gene detection.In addition,97 specimens were tested for EML4-ALK fusion gene alterations using the Ventana method,and the results were compared with that by PCR detection.The mutation characteristics were summarized,and the cytological features of the mutations and fusions were observed.Results The positive rate of driver gene mutations in 49 cases of multi-gene panel detection was 65.31%,which indicated no statistically significant difference from that of 99 lung adenocarcinoma tissue specimens(P>0.05).The positive rates of ALK,ROS1,and RET fusion mutations were 4.08%,4.08%,and 2.04%,respectively,and the positive rates of EGFR,KRAS,HER2,MET,and BRAF mutations were 38.78%,6.12%,4.08%,4.08%,and 2.04%,respectively.The mutation rate of EGFR in 151 cases of single-gene detection was 54.97%,with a mutation rate of 42.86%in males and 67.57%in females,showing a statistically significant difference between genders(P<0.05).Exon 19 deletion mutation(19-Del)and exon 21 point mutation(21-L858R)in the EGFR gene took the highest proportions,accounting for 54.22%and 37.35%,respectively,significantly higher than other mutation types(P<0.001).The positive rate of EML4-ALK fusion gene immunohistochemistry(IHC)in 97 cytological paraffin-embedded specimens was 15.46%,among which 6 cases with weak positivity(+)showed negative results in molecular detection,and 9 cases with diffuse positivity()showed positive results in molecular detection.The results of immunohistochemistry were highly consistent with molecular detection in cases with diffuse strong positivity(Kappa=1).It was found from the cytological morphology of cases with gene alterations that KARS mu

关 键 词：肺腺癌 细胞蜡块 驱动基因 联合检测 免疫组织化学 靶向治疗 

分 类 号：R734.2[医药卫生—肿瘤]