非小细胞肺癌患者肿瘤组织多驱动基因联合检测及其临床意义

目的探讨EGFR、KRAS基因突变及ALK、ROS1基因融合在非小细胞肺癌(non-small cell lung cancer,NSCLC)患者中的检出率,并分析与NSCLC临床病理特征的关系。方法收集NSCLC手术标本86例,采用荧光PCR法检测EGFR、KRAS突变及ALK、ROS1基因融合,并分析EGFR、KRAS、ALK及ROS1基因改变与患者性别、年龄、吸烟史、组织学类型、有无淋巴结转移等临床病理特征的相关性。结果 NSCLC肿瘤组织中驱动基因总突变率为62.8%(54/86),其中EGFR基因突变占总突变的76.0%(41/54);KRAS基因突变占总突变的9.3%(5/54);ALK基因融合占总突变的13.0%(7/54),其中1例患者存在EGFR 19缺失突变与ALK融合共存;ROS1基因融合占总突变的3.8%(2/54)。NSCLC的临床病理特征显示,EGFR基因突变在女性、腺癌患者中突变率高(P0.05);与患者年龄、是否吸烟、有无淋巴结转移无明显相关(P0.05);KRAS、ALK、ROS1基因改变与NSCLC的临床病理特征无明显相关(P0.05)。结论 NSCLC中EGFR、ALK基因均存在较高的突变率,临床医师应给予高度重视;KRAS、ROS1基因改变以及驱动基因双突变共存型基因突变率虽低,但其意义重大不容忽视。     

非小细胞肺癌表皮生长因子受体、间变性淋巴瘤激酶、ROS1基因突变及突变共存的临床病理学意义

目的探讨非小细胞肺癌表皮生长因子受体(EGFR)基因突变、间变性淋巴瘤激酶(ALK)及ROS1基因融合突变情况与临床病理特征的关系,并分析驱动基因共突变的病例。方法收集福建医科大学附属协和医院1508例非小细胞肺癌患者临床病理资料,采用荧光PCR法检测EGFR基因突变及ALK、ROS1基因融合突变情况,统计分析驱动基因突变状态与临床病理特征的关系。结果1508例非小细胞肺癌中,EGFR、ALK、ROS1基因突变率分别为52.9%(797/1508)、6.2%(93/1508)、2.7%(40/1508);EGFR基因突变类型以外显子19 del与外显子21 L858R为主(90.6%,722/797),EGFR基因突变多见于女性、无吸烟史、腺癌患者(P<0.05);ALK及ROS1基因融合多见于年龄<60岁、晚期患者(P<0.05)。其中携带共突变基因的非小细胞肺癌患者共16例(1.1%,16/1508),包括EGFR/ALK基因共突变7例、EGFR/ROS1基因共突变8例、ALK/ROS1基因共突变1例,驱动基因共突变患者多为女性、腺癌患者。16例驱动基因共突变患者中,8例使用酪氨酸激酶抑制剂治疗,疗效为3例部分缓解、2例疾病稳定、3例疾病进展。结论非小细胞肺癌驱动基因可出现突变共存的状态,多基因联合检测可为临床靶向药物治疗策略的制定提供参考意义。     

基于二代测序的300例非小细胞肺癌中驱动基因突变与临床病理特征的关系

目的探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)常见驱动基因的突变特点及其与临床病理特征的关系。方法采用二代测序技术(next generation sequencing,NGS)检测300例NSCLC石蜡包埋组织中常见驱动基因如EGFR、KRAS、ALK、ROS1、BRAF、MET、RET、HER-2等的突变情况。结果 300例NSCLC中,EGFR、KRAS、ALK、ROS1、BRAF、MET、RET、HER-2等基因的突变率分别为52. 00%、10. 33%、6. 67%、1. 67%、3. 67%、3. 33%、1. 00%、2. 33%。EGFR 21外显子L858R突变合并LINCO1446-EGFR基因融合1例; EGFR 20外显子C797S与T790M呈顺式或反式存在并合并EGFR敏感突变各1例; EGFR基因点突变合并MET基因拷贝数扩增3例。EGFR突变多见于不吸烟的女性肺腺癌患者(P 0. 05); KRAS突变多见于吸烟男性患者(P 0. 05); ALK突变与患者年龄有关(P 0. 05),在小于60岁患者中更常见; ROS1融合突变与患者性别有关(P 0. 05),女性多见; BRAF、MET、RET、HER-2基因突变与患者性别、年龄、吸烟、组织学类型及c TNM分期无关。结论 EGFR可与其他驱动基因突变共存;基因突变与患者性别、年龄、吸烟与否、组织学类型有相关性; BRAF、MET、RET、HER-2的突变率低,其临床意义有待探讨; NGS发现的共存型基因突变及少见突变应引起重视。     

非小细胞肺癌患者肿瘤组织中驱动基因的分子病理检测分析

目的：探讨非小细胞肺癌患者肿瘤组织中驱动基因EGFR、KRAS、ALK、ROS1、c-Met和Her-2基因各亚型改变情况。方法：应用Taqman-ARMS方法检测273例非小细胞肺癌石蜡组织中EGFR基因、KRAS基因、ALK基因、ROS1基因、c-Met基因和Her-2基因改变情况。结果：非小细胞肺癌肿瘤组织中EGFR基因总突变率为36.26%(99/273),外显子18、19、20和21的突变率分别为0(0/273)、16.12%(44/273)、4.49%(15/273)和17.95%(49/273);EGFR基因各外显子之间双重突变共12例(4.40%);KRAS基因总突变率为4.76%(13/273);ALK融合基因总阳性率为9.16%(25/273);ROS1融合基因总阳性率为2.20%(6/273);c-Met基因总扩增率为3.66%(10/273);Her-2基因总突变率为0.73%(2/273);各驱动基因双突变共存型11例(4.03%),其中EGFR基因突变与ALK融合基因阳性共存型2例(0.73%),EGFR基因突变与KRAS基因突变共存型3例(1.10%),EGFR基因突变与c-Met基因扩增共存型6例(2.20%)。结论：非小细胞肺癌患者中EGFR基因19和21外显子突变和ALK融合基因均存在较高的突变率,基因突变亚型分类能指导精准医学的个体化靶向治疗,KRAS、ROS1、c-Met、Her-2基因改变以及驱动基因双突变共存型基因突变率虽低但不容忽视。     

非小细胞肺癌中九基因联合检测突变分析

目的通过ARMS-PCR法联合检测非小细胞肺癌(non-small cell lung cancers, NSCLC)中9个驱动基因(ALK、ROS1、RET、EGFR、KRAS、HER-2、PIK3CA、NRAS和BRAF)的突变情况,分析其突变状态及临床意义。方法采用ARMS-PCR技术检测2018年2月～2019年2月陆军军医大学第一附属医院病理科存档的522例NSCLC肿瘤组织中的9个驱动基因的突变情况。结果 522例NSCLC中ALK、ROS1和RET的融合突变率分别为5.17%、1.34%、1.34%,EGFR、KRAS、HER-2、PIK3CA、NRAS和BRAF的突变率分别为47.32%、7.28%、1.72%、1.72%、0.95%和0.57%。女性患者中EGFR突变和ALK融合突变率明显高于男性患者(P0.001),而KRAS突变率低于男性患者(P0.001)。EFGR和KRAS突变在肺腺癌中显著高于肺鳞癌(P0.001)。无吸烟史患者中EGFR突变和ALK融合发生率均高于吸烟患者(P0.001),KRAS突变在吸烟患者的发生率显著高于无吸烟史患者。利用ARMS法联合检测9个基因在单点检测EGFR基础上增加了10.15%的患者可使用靶向药物(P0.01)。结论在9个驱动基因突变中,EGFR突变、ALK融合、KRAS突变与患者性别、吸烟以及组织学类型密切相关,其它较为罕见驱动基因突变并未发现与组织学类型、患者性别及吸烟与否相关。九基因联合检测可作为NSCLC更简便适用的药物靶向检测方法。     

肺腺癌基因谱分析及p53、KRAS的表达及意义

目的分析肺腺癌基因谱,探讨p53和KRAS在肺腺癌中的表达及其意义。方法采用二代测序技术检测29例肺腺癌石蜡样本中26个常见肿瘤相关热点基因;采用免疫组化法检测99例肺腺癌组织中p53蛋白和KRAS蛋白的表达,分析两者表达与肺腺癌临床病理特征的关系。结果 26个被检基因中发现6个基因发生突变,分别是EGFR(21/29)、p53(12/29)、KRAS(3/29)、ROS1(3/29)、ALK(1/29)和RET(1/29);EGFR突变多发生在19号(9/21例)和21号(10/21例)外显子上,2例发生在20号外显子上(其中1例在20和21号外显子上均有突变),1例发生在18号外显子上;p53突变位于6号(4/12)、4号(2/12)、5号(2/12)和8号(2/12)、7号(1/12)和10号(1/12)外显子上;KRAS突变均发生在2号(3/3)外显子上。29例中14例存在共突变,其中1例发生3种基因共突变(EGFR、KRAS、ROS1),13例发生双基因突变(10例EGFR和p53,1例EGFR和KRAS、1例EGFR和ROS1,1例p53和KRAS),12例发生单基因突变,3例未检出基因突变。免疫组化检测结果显示肺腺癌组织中p53阳性率为30.3%,KRAS阳性率为23.2%,两者表达无明显相关(r=0.054,P=0.594)。肺腺癌中p53表达与患者性别、年龄、病理亚型、肿瘤大小、有无淋巴结转移、TNM分期和分化程度均无关(P0.05),KRAS表达也与患者年龄、病理亚型、肿瘤大小、有无淋巴结转移、TNM分期和分化程度无关(P0.05),但KRAS在男性(36.4%)肺腺癌中的表达高于女性(12.7%)(P0.05)。结论肺腺癌组织中存在多种基因突变,并可发现共突变,EGFR突变多在19和21号外显子上,p53突变多集中在6、4、5和8号外显子上,KRAS突变多在2号外显子上,且KRAS突变更多见于男性患者。     

EGFR及KRAS基因突变与非小细胞肺癌临床病理特征的关系

目的探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)患者EGFR及KRAS基因突变与其临床病理特征的关系。方法采用毛细管电泳法及荧光探针法分别检测64例NSCLC组织中EGFR及KRAS基因的突变类型。结果 64例NSCLC中发生EGFR基因突变27例(占42.2%)、KRAS基因突变8例(占12.5%),同时发生EGFR和KRAS基因突变者4例(占6.25%)。EGFR基因突变与患者性别、组织学类型及吸烟史有关(P0.05),与患者年龄、分化程度、有无淋巴结转移及TNM分期无关(P0.05)。肺腺癌中KRAS基因突变率明显高于肺鳞癌(P0.01),KRAS基因突变与患者性别、年龄、有无吸烟史、分化程度、有无淋巴结转移及TNM分期均无关(P0.05)。结论 NSCLC患者中EGFR基因突变率高于KRAS基因突变率,EGFR突变率在女性、肺腺癌、不吸烟患者中较高,KRAS基因突变率在腺癌患者中较高,且EGFR和KRAS基因突变可以同时发生。     

肺腺癌EGFR基因19、21号外显子突变分析

目的 探讨肺腺癌表皮生长因子受体(EGFR)基因19、21号外显子突变特点及其临床意义.方法 对肺腺癌手术组织提取DNA,采用聚合酶链式反应(PCR)和直接测序法检测EGFR基因19号与21号外显子突变状态,分析与临床病理参数之间的相关性.结果 在54例被测肺腺癌肿瘤中,发现29例患者存在EGFR基因突变,占53.7％ (29/54).其中,19号外显子突变13例(24.1％),均为缺失突变;21号外显子突变病例为16例(29.6％),均为L858R点突变.高、中分化肿瘤的EGFR突变率均达60％以上,明显高于低分化肿瘤突变率44％.结论 肺腺癌存在较高频率的EGFR基因突变,在高、中分化肿瘤的突变率有高于低分化肿瘤的趋势.     

非小细胞肺癌中EGFR、ALK、KRAS基因突变及PD-L1表达的关系

目的探讨非小细胞肺癌(non-small cell lung cancer, NSCLC)中EGFR、KRAS、ALK基因突变状态及与PD-L1蛋白表达的关系。方法收集中国科学技术大学附属第一医院2019年9月30日～2020年9月30日收治的128例NSCLC,采用免疫组化法及二代测序(next generation sequencing, NGS)技术对其PD-L1表达、EGFR、ALK、KRAS基因突变状态进行检测,结合患者临床资料,对PD-L1蛋白表达、EGFR、KRAS、ALK基因突变的临床病理特征及相关性进行分析。结果 NSCLC中,女性、非吸烟、肺腺癌患者的EGFR突变率较高(P0.05);ALK融合在女性患者中阳性率高,且更易发生胸膜转移(P0.05);PD-L1蛋白阳性率为69.53%,男性(P=0.073)、非吸烟患者(P=0.022)的PD-L1阳性率相对较高。相关性分析显示,PD-L1表达与KRAS基因突变呈正相关(r=0.203,P=0.022),与EGFR基因突变呈负相关(r=-0.149,P=0.093),与ALK基因突变无相关性(r=0.067,P0.05)。结论 NSCLC中PD-L1表达与EGFR、KRAS基因突变有相关性,需要进一步研究其对临床免疫治疗相关的预测意义。     

针吸活检与基因突变检测诊断非小细胞肺癌的临床价值

目的探讨经支气管镜针吸活检(Transbronchial needle aspiration,TBNA)组织学和基因突变检测在非小细胞肺癌的分型诊断及治疗中的价值。方法探讨经支气管镜针吸活检组织学和基因突变检测在非小细胞肺癌的分型诊断及治疗中的价值。结果进行TTF-1、p63、ck7、ck5/6特异性标记物免疫组化检测的28例TBNA样本的检测结果为:鳞癌15例,腺癌13例。其中10例腺癌进行了EGFR/KRAS/ALK的基因突变检测,检出EGFR突变阳性2例,KRAS阳性1例,未检测到ALK突变。结论通过TTF-1、p63、ck7、ck5/6特异性标记物联合基因突变检测可以较准确区分出鳞癌和腺癌,为临床用药提供指导。     

276例手术切除肺腺癌亚型与EGFR/ALK基因状态

目的:探讨手术切除肺腺癌各亚型EGFR和ALK基因状态分布.方法:应用ARMS方法检测手术切除肺腺癌石蜡组织中EGFR基因突变和ALK融合基因情况.结果:276例肺腺癌手术样本中,EGFR基因突变率为54.71％(151/276),其中19del为28.99％(80/276),L858R为23.19％(64/276),20-ins为0.72％(2/276),L861Q为0.72％(2/276),G719X为1.09％(3/276),S768I为0.36％(1/276)和T790M为0.72％(2/276),其中包含G719X+S768I,19del+T790M,L858R+T790M各1例,ALK基因融合阳性率为5.80％(12/207),在肺腺癌各亚型中EGFR基因突变附壁状腺癌,腺泡状腺癌,乳头状腺癌,实体状腺癌和浸润性黏液腺癌之间差异有统计学意义(P＜0.001,P=0.009,P=0.023,P＜0.001和P=0.030),与其他类型之间差异均无统计学意义(P＞0.05);在肺腺癌各亚型中ALK融合基因突变各亚型之间差异均无统计学意义(P＞0.05).结论:肺腺癌组织学亚型与EGFR基因突变有关,附壁状腺癌、腺泡状腺癌和乳头状腺癌出现EGFR基因突变比其他亚型更加明显.     

肺鳞状细胞癌EGFR/ALK的基因状态

目的:探讨肺鳞状细胞癌各亚型中EGFR和ALK的基因状态.方法:应用ARMS方法检测肺鳞状细胞癌石蜡组织中EGFR基因突变和ALK融合基因情况.结果:218例肺鳞状细胞癌样本中,E GFR基因突变率为4.59％(10/218),19del和L858R各为2.29％(5/218).ALK融合基因阳性率为6.14％(7/114).结论:肺鳞状细胞癌存在一定比例EGFR基因突变和ALK融合基因阳性,肺鳞状细胞癌EGFR基因和ALK融合基因常规检测不可忽视.     

非小细胞肺癌患者肿瘤组织中EGFR和KARS基因突变的分子病理检测分析

目的：探讨非小细胞肺癌患者肿瘤组织中EGFR和KRAS基因各亚型突变情况。方法：应用直接测序方法检测非小细胞肺癌石蜡组织中1273例EGFR基因和1062例KRAS基因突变情况。结果：非小细胞肺癌肿瘤组织中EGFR基因总突变率为36.68%(467/1273),外显子18、19、20和21的突变率分别为1.02%(13/1273)、18.93%(241/1273)、2.59%(33/1273)和15.95%(203/1273);EGFR基因各外显子之间双重突变共17例(1.34%),其中18外显子与20外显子双重突变3例(0.24%),19外显子与20外显子双重突变7例(0.55%),19外显子与21外显子双重突变4例(0.31%)和20外显子与21外显子双重突变3例(0.24%);EGFR基因各外显子内双重突变共2例(2.18%),均为21外显子双重突变。KRAS基因总突变率为3.01%(32/1062),外显子2的密码子5、12、13和25的突变率分别为0.09%(1/1062)、2.64%(28/1062)、0.18%(2/1062)和0.09%(1/1062),外显子3密码子61的突变率为0.09%(1/1062)。结论：非小细胞肺癌患者中EGFR基因存在较高的突变率,尤其为19和21外显子突变,其基因突变亚型分类能指导EGFR-TKI的肿瘤靶向治疗,KRAS基因突变率虽低但不容忽视,其基因突变预示着EGFR-TKI原发耐药。     

Analysis of multigene detection in patients with advanced lung adenocarcinoma using cytological specimens

ObjectiveTo investigate the mutation status and clinical characteristics of multigene detection in advanced lung adenocarcinoma using cytological specimens.Materials and Methods137 advanced lung adenocarcinoma patients with 10 driver genes detection in the Fourth Hospital Hebei Medical University from January 2019 to November 2019 was analysized. 137 cytological specimens including fine-needle aspiration specimens and maligant serous cavity effusion (pleural effusion, peritoneal and pericardial effusion). Ten driver mutations of EGFR, ALK, ROS1, BRAF, KRAS, NRAS, HER2, RET, PIK3CA and MET were detected by the amplification refractory mutation system (ARMS). Meanwhile, 90 of 137 patients were detected with biopsies for parallel gene detection.Results78.10 % (107/137) of patients with advanced lung adenocarcinoma harbored at least one of 10 driver mutations. The three main mutations were EGFR (69.16 %, 74/137), ALK (6.57 %, 9/137)and ROS1 (3.65 %, 5/137) mutations. Besides, we found 6 cases including two concomitant mutations: EGFR Exon19 del/HER2 (1/137), EGFR Exon21 L858R/PIK3CA (2/137), EGFR Exon21 L858R/RET (1/137), and ALK/KRAS (2/137). Among 137 patients, women aged 64 or older were more likely to have the mutations (P < 0.05). Female patients (P = 0.003) older or equal to 64 years (P = 0.015) with non-smoking habbit (P = 0.027) were more detected with EGFR mutations, while ALK was more detectable in patients yonger than 64 years. Parallel analysis showed that rates of single EGFR, ALK, ROS1, RET, KRAS, NRAS, HER2, MET mutations and concomitant different mutations were not significantly different between cytological specimens and matched histological specimens.ConclusionsIn the study, cytological specimens and biopsy samples have a very high coincidence rate of gene detection. EGFR, ALK and ROS1 mutations were the main driver mutations in patients with advanced lung adenocarcinoma.We speculate that EGFR and ALK are more prone to concomitant mutations respectively and the treatment of advanced lung adenocarcinoma patients with concomitant mutations deserves further study. The rate of KRAS, NRAS, BRAF, PIK3CA, RET and MET exon14 skipping mutation were low but may had a significant impact on the targeted therapy of patients with advanced lung adenocarcinoma.     

EGFR突变的非小细胞肺癌患者EML4-ALK融合基因的检测及其临床特征分析

目的: 检测EML4-ALK融合基因在表皮生长因子受体(epidermal growth factor receptor, EGFR)突变的非小细胞肺癌(non-small-cell lung cancer, NSCLC)人群中的突变率,并分析其与临床特征的关系。方法: 入选的102例NSCLC患者均为中国人,且至少满足以下1个入选条件:女性、不吸烟/少吸烟和肺腺癌。将102例患者的组织标本采用多重逆转录聚合酶链反应 (multiplex RT-PCR)的方法检测其EML4-ALK融合基因的突变率;对EML4-ALK阳性患者的组织标本采用DNA扩增后直接测序的方法来检测其EGFR(18~21号外显子)及Kirsten鼠肉瘤基因(Kirsten rat sarcoma,KRAS)(1、2号外显子)的突变情况。结果: 102例非小细胞肺癌患者的组织标本,有8例(7.8%)存在EML4-ALK融合基因突变,其中7例为突变体1(variant 1,V1),1例为突变体2(variant 2,V2);这8例EML4-ALK阳性患者组织标本的EGFR(18~21号外显子)及KRAS(1、2号外显子)均为野生型。8例阳性患者中,5例患者的年龄小于总体患者的平均年龄(59±10)岁,占62.5%(5/8);女性患者6例,占75%(6/8);不吸烟患者7例,占87.5%(7/8);腺癌患者5例,占62.5%(5/8)。结论: EML4-ALK融合基因突变代表了NSCLC的一个新的分子亚型,EML4-ALK突变与EGFR及KRAS突变是不共存的。     

非小细胞肺癌表皮生长因子受体基因突变与临床病理特征的关系

目的探讨非小细胞肺癌(NSCLC)表皮生长因子受体(EGFR)基因突变的临床病理学意义。方法应用即时荧光定量聚合酶链反应(PCR)法检测1444例NSCLC患者肺癌组织中EGFR基因第19号和21号外显子突变状况,分析EGFR基因突变与年龄、性别、吸烟状况、组织学分级及临床分期之间的关系。结果1410例成功分型的NSCLC肿瘤组织中401例存在EGFR突变,突变检出率为27.8％,其中193例第19号外显子发生缺失突变,208例第21号外显子发生替代突变。EGFR突变更常见于腺癌、不吸烟或女性患者[突变率分别为33.5％ (367/1094)]、37.6％( 321/853)及43.2％ (225/521)。细支气管肺泡癌及伴细支气管肺泡癌分化特征的腺癌突变率显著高于普通腺癌[突变率分别为61.3％( 19/31)、48.0％( 12/25)及32.4％ (336/1038)]。随着吸烟暴露水平的增加,EGFR突变率呈下降趋势;女性、不吸烟、腺癌为EGFR基因突变状况的独立影响因素。结论NSCLC患者女性、不吸烟、腺癌患者突变率较高。即时荧光定量PCR技术可快速、敏感、准确地检测EGFR基因突变。     

非小细胞肺癌组织表皮生长因子受体基因第19外显子突变研究

目的 研究非小细胞肺癌的表皮生长因子受体(epidermal growth factor receptor,EGFR)基因第19外显子突变情况.方法 提取72例手术切除肺癌基冈组DNA,用巢式PCR扩增EGFR基因第19外显子并测序,分析EGFR基因第19外显子突变及其与I临床特征的相关性.结果 检测到13例肿瘤组织存在EGFR基因第19外显子多核苷酸的缺失突变,共有4种突变类型,突变率为18.1%,所检测的突变均为杂合型;EGFR基因第19外显子突变与肺腺癌、女性患者与不吸烟者相关.结论 EGFR基因第19外显子存在多核苷酸的框架缺失突变,以女性、腺癌和不吸烟患者为主.     

EGFR, ERBB2, and KRAS mutations in Korean non-small cell lung cancer patients

The epidermal growth factor receptor (EGFR), and its family members play an important role in the development and progression of lung cancers. It has been reported that somatic mutations in the tyrosine kinase domain of the EGFR or ERBB2 genes occur in a subset of patients with lung cancer. We searched for mutations of the EGFR, ERBB2, and KRAS genes in surgically resected non-small cell lung cancers (NSCLCs) to determine the prevalence of these mutations in Korean lung cancer patients. In addition, we examined the relationship between the mutations and clinicopathologic features of lung cancers. Mutations of the EGFR, ERBB2, and KRAS genes were determined by polymerase chain reaction-based direct sequencing in 115 surgically resected non-small cell lung cancers. EGFR mutations were present in 20 patients (17.4%). The EGFR mutations were found only in adenocarcinomas (20 of 55 adenocarcinomas, 36.4%). The ERBB2 mutation was found in 1 adenocarcinoma of the 115 NSCLCs (0.9% overall; 1.8% of the 55 adenocarcinomas). KRAS mutations were found in 6 (5.2%) of the 115 NSCLCs (2 of 60 squamous cell carcinomas, or 3.3%, and 4 of 55 adenocarcinomas, or 7.3%). EGFR mutations in adenocarcinomas were more frequent in women (P = 0.02) and in never-smokers (P = 0.004). EGFR mutations in adenocarcinomas were not associated with pathologic stage in never-smokers, but were more frequent in pathologic stage II-IV than in stage I in ever-smokers (P = 0.01). Of the 55 adenocarcinomas, 25 (45.5%) had mutations of one or another of the three genes; EGFR mutations were never found in adenocarcinomas together with ERBB2 or KRAS mutations. These findings suggest that the EGFR mutation is frequent in Korean lung cancer patients, and that the ERBB2 mutation is rare. Further studies are needed to investigate the role of EGFR mutations in the carcinogenesis of adenocarcinoma among smokers.     

肺癌表皮生长因子受体基因突变和扩增与临床病理相关性研究

目的探讨肺癌患者中表皮生长因子受体(EGFR)基因突变和扩增与临床病理的关系。方法应用聚合酶链反应(PCR)-DNA测序和荧光原位杂交(FISH)检测肺癌EGFR基因突变和扩增的情况,并结合临床病理资料进行分析。结果在DNA测序检测的454例肺癌中,总突变率48.2％ (219/454)。女性患者突变率59.6％ (118/198)明显高于男性39.5％ (101/256),两者差异有统计学意义(P＜0.001)。非吸烟者突变率52.7％( 147/279)高于吸烟者41.1％ (72/175),两者差异有统计学意义(P=0.017)。腺癌突变率56.8％ (193/340)高于非腺癌者22.8％ (26/114),两者之间差异有统计学意义(P＜0.05)。腺癌各亚型之间突变率差异有统计学意义(P=0.001)。在FISH检测的134例肺癌中,基因扩增阳性率22.4％ (30/134),其在年龄、性别、组织学类型和腺癌亚型分型中无统计学意义(P＞0.05)。EGFR基因扩增与基因突变之间具有显著相关性（P=0.0005）,但两者一致性较差(κ=0.275)。结论EGFR基因在女性、非吸烟者和腺癌患者中突变率高,且腺癌各亚型之间突变率有明显差异;EGFR基因扩增与年龄、性别、组织学类型和腺癌亚型分型无明显相关。DNA测序与FISH两种检测方法之间具有显著相关性,但两者一致性较差。     

Molecular diagnostic characteristics based on the next generation sequencing in lung cancer and its relationship with the expression of PD-L1

BackgroundNext generation sequencing (NGS) is a massively parallel sequencing technique that can be used to detect many forms of DNA variation, including point mutations, small fragment insertion deletions, gene recombination, and copy number variations. It can simultaneously analyze multiple genes and mutations, quantitatively detect gene mutation rate, and provide comprehensive information for clinicians. More and more lung cancer patients have benefited from studies on programmed death-1igand l (PD-L1) and immunocheckpoint inhibitors. The relationship between gene mutation and PD-L1 is also a focus of current research. Therefore, we collected a large number of cases to describe the molecular diagnostic characteristics of NGS in lung cancer and the relationship between NGS and PD-L1 expression.MethodA total of 1017 lung cancer patients with 15-gene panel (EGFR, ALK, ROS1, BRAF, MET, RET, ERBB2, KRAS, PIK3CA, KIT, ESR1, PDGFRA, DDR2, HRAS, NRAS) examined by NGS from our hospital were collected to analyze their clinicopathological characteristics. 600 of 1017 patients were tested for PD-L1 (22C3) by immunohistochemistry (IHC) at the same time. PD-L1 tumor proportion score (TPS) were used for comparative analysis with gene mutation results, and then to screen for possible correlation genes.Results74.63 % (759/1017) of lung cancer patients had at least one version of the genes. The top three mutation were EGFR (46.41 %), KRAS (13.86 %) and PIK3CA (10.03 %). Mutations in EGFR, KRAS, PIK3CA, KIT, ESR1 and NRAS were associated with sex (P < 0.05). Except for EGFR, which was more frequent in female, other genes were more frequent in male. ALK was more detectable in patients younger than 60, while PIK3CA was more detectable in patients older than 60(P < 0.05). EGFR, ALK, ROS1, KRAS, PIK3CA, ESR1 and NRAS were associated with smoking (P < 0.05). EGFR, KRAS, PIK3CA and ESR1 were correlated with pathological histology (P < 0.05). Among the 15 genes, only EGFR was associated with pathological histology of invasive adenocarcinoma (IA). EGFR had the highest mutation rate (60.00 %) in Lepidic predominate IA. Significantly different in sample types were found in EGFR, ALK, MET, KRAS, PIK3CA and NRAS examined by NGS. There were significant differences in the TPS of PD-L1 (22C3) in EGFR, ALK, BRAF and MET variants (P < 0.05). EGFR mutations were more common in TPS < 1 %, ALK mutations were more common in TPS (1 %–49 %), and BRAF and MET mutations were more common in TPS ≥ 50 %.ConclusionIn the 15-gene panel, in addition to EGFR, ALK and ROS1, MET, KRAS, PIK3CA, KIT, ESR1 and NRAS also had their own characteristics in sex, age, smoking history, histopathology, sample type and PD-L1, showing different clinicopathological tendencies. Understanding this information can help us optimize stratified lung cancer patients. Furthermore, it provides patients with a variety of diagnostic needs and a large number of unique clinical data worthy of clinical recognition.