학술논문
非小细胞肺癌中表皮生长因子受体基因突变直接测序分析及其与临床病理特征的相关性 / Detection of epidermal growth factor receptor mutations in non-small-cell lung carcinoma by direct sequencing and correlations with clinicopathological characteristics and sample types
Document Type
Academic Journal
Author
孙孟红; 杨飞; 沈磊; 张玲; 陈颖; 蔡旭; 朱晓丽; 周晓燕; SUN Meng-hong; YANG Fei; SHEN Lei; ZHANG Ling; CHEN Ying; CAI Xu; ZHU Xiao-li; ZHOU Xiao-yan
Source
中华病理学杂志 / Chinese Journal of Pathology. 40(10):655-659
Subject
Language
Chinese
ISSN
0529-5807
Abstract
目的回顾性分析非小细胞肺癌(NSCLC)中采用直接测序法检测的表皮生长因子受体(EGFR)基因突变率、突变分布特征及其与临床病理的相关性。方法收集NSCLC共443例，其中包括手术切除样本299例、粗针穿刺活检标本59例、细针穿刺和胸腔积液细胞学样本85例。所有样本均通过石蜡包埋、切片后确定肿瘤细胞含量，显微切割富集肿瘤细胞后采用聚合酶链反应-直接测序法检测EGFR基因酪氨酸激酶编码区第18至21号外显子的突变。结果(1)在443例NSCLC标本中共检出EGFR基因突变193个，发生在189例患者中(42.7％)；EGFR基因第18至21号外显子的突变率分别为2.0％( 4/193)、48.7％ (94/193)、6.7％ (13/193)和42.5％ (82/193)；(2)EGFR基因总突变率与年龄无显著相关性，但在年龄大于中位年龄（57岁）的患者中第21号外显子的突变率(50.9％，54/106)高于年龄小于或等于中位年龄者(32.2％，28/87；P＜0.01)；(3)女性患者中EGFR基因突变率(53.5％,107/200)高于男性患者突变率(33.7％，82/243；P ＜0.01)；(4)腺癌中的突变率(46.5％，161/346)高于鳞癌(13.3％，4/30；P＜0.01)和低分化癌者(24.1％，7/29；P＜0.05)，而与腺鳞癌(7/13)差异无统计学意义(P＞0.05)；(5)在手术切除、粗针穿刺活检以及细针穿刺和胸腔积液细胞学样本中EGFR突变检出率有逐渐下降的趋势，分别为49.5％( 148/299)、35.6％ (21/59)和23.5％ (20/85)，其中细针穿刺和胸腔积液的细胞学样本中的检出率较手术标本中的检出率为低(P＜0.01)。结论直接测序法是检测NSCLC中EGFR基因突变的有效方法，特别对未知突变类型的检出具有优势；NSCLC中EGFR基因突变在女性和腺癌中多见，以第19号外显子的缺失突变和第21号外显子的点突变为主；EGFR基因突变分布特征可能与年龄有一定关系；EGFR基因突变检出率与样本类型密切相关，活检组织及细胞学样本是检测EGFR基因突变的有用材料，但可能会漏检部分患者中EGFR基因的突变。
Objective To retrospectively analyze epidermal growth factor receptor (EGFR) gene mutation frequencies and distribution characteristics in Chinese patients with non-small-cell lung carcinoma (NSCLC) by direct gene sequencing. Methods Clinical samples from 443 NSCLC patients were obtained for EGFR gene mutation analysis, including 299 surgical specimens, 59 core biopsies and 85 fine needle aspiration and pleural effusion cytology specimens. All samples were processed from paraffin embedded blocks and microdissection was performed to enrich tumor cells. PCR based direct gene sequencing was used to investigate tyrosine kinase domain coding region involving exon 18 through 21. Results ( 1 ) Among 443 samples, 193 mutations were detected in 189 patients (42.7％ ) and 4 patients possessed two mutations involving two different exons in their tumor samples. The percentage of mutations involving exon 18 to 21 were 2.0％ (4/193) ,48.7％ (94/193) ,6.7％ (13/193) and 42.5％ (82/193) respectively. (2) There was no significant correlation of EGFR mutation with age, however, mutation rate (50.9％, 54/106) of exon 21 in patients over median age 57 was higher than that of the younger patients (32.2％ ,28/87;P ＜0.01 ). (3) EGFR mutation rate was remarkably higher in female patients (53.5％ ,107/200) than in male patients (33.7％ ,82/243 ; P ＜ 0.01 ). ( 4 ) Mutation rate in adenocarcinomas (46.5％, 161/346 ) was much higher than in squamous cell carcinomas (13.3％, 4/30) and poorly differentiated carcinomas (24.1％ ,7/29 ;P ＜ 0.01, P ＜0.05 ), while the adenosquamous carcinomas shared a mutation rate similar to that of adenocarcinoma (7/13, P ＞0.05). (5) In surgical samples, core biopsies and cytological samples, the EGFR mutation detection rates were 49.5％ (148/299) ,35.6％ (21/59)and 23.5％ (20/85)respectively. The fine needle aspiration and cytological samples showed much lower EGFR mutation detection rates (23.5％ ,20/85 ) than that of surgical samples (49.5％, 148/299 ; P ＜ 0.01 ). Conclusions ( 1 )Direct gene sequencing is a reliable and effective method for the detection of EGFR mutations in NSCLC,particularly for unknown EGFR mutations. (2) EGFR mutations are more frequent in female patients and patients with adenocarcinoma NSCLC, involving mainly exon 19 and 21. (3) The mutation distribution in exons of EGFR gene appears age-related. (4) Detection rate of EGFR mutation varies in different sample types. Small biopsy and fine needle aspiration specimens are valuable materials for analyzing EGFR mutation in NSCLC, although rare false negativity may occur using such samples.
Objective To retrospectively analyze epidermal growth factor receptor (EGFR) gene mutation frequencies and distribution characteristics in Chinese patients with non-small-cell lung carcinoma (NSCLC) by direct gene sequencing. Methods Clinical samples from 443 NSCLC patients were obtained for EGFR gene mutation analysis, including 299 surgical specimens, 59 core biopsies and 85 fine needle aspiration and pleural effusion cytology specimens. All samples were processed from paraffin embedded blocks and microdissection was performed to enrich tumor cells. PCR based direct gene sequencing was used to investigate tyrosine kinase domain coding region involving exon 18 through 21. Results ( 1 ) Among 443 samples, 193 mutations were detected in 189 patients (42.7％ ) and 4 patients possessed two mutations involving two different exons in their tumor samples. The percentage of mutations involving exon 18 to 21 were 2.0％ (4/193) ,48.7％ (94/193) ,6.7％ (13/193) and 42.5％ (82/193) respectively. (2) There was no significant correlation of EGFR mutation with age, however, mutation rate (50.9％, 54/106) of exon 21 in patients over median age 57 was higher than that of the younger patients (32.2％ ,28/87;P ＜0.01 ). (3) EGFR mutation rate was remarkably higher in female patients (53.5％ ,107/200) than in male patients (33.7％ ,82/243 ; P ＜ 0.01 ). ( 4 ) Mutation rate in adenocarcinomas (46.5％, 161/346 ) was much higher than in squamous cell carcinomas (13.3％, 4/30) and poorly differentiated carcinomas (24.1％ ,7/29 ;P ＜ 0.01, P ＜0.05 ), while the adenosquamous carcinomas shared a mutation rate similar to that of adenocarcinoma (7/13, P ＞0.05). (5) In surgical samples, core biopsies and cytological samples, the EGFR mutation detection rates were 49.5％ (148/299) ,35.6％ (21/59)and 23.5％ (20/85)respectively. The fine needle aspiration and cytological samples showed much lower EGFR mutation detection rates (23.5％ ,20/85 ) than that of surgical samples (49.5％, 148/299 ; P ＜ 0.01 ). Conclusions ( 1 )Direct gene sequencing is a reliable and effective method for the detection of EGFR mutations in NSCLC,particularly for unknown EGFR mutations. (2) EGFR mutations are more frequent in female patients and patients with adenocarcinoma NSCLC, involving mainly exon 19 and 21. (3) The mutation distribution in exons of EGFR gene appears age-related. (4) Detection rate of EGFR mutation varies in different sample types. Small biopsy and fine needle aspiration specimens are valuable materials for analyzing EGFR mutation in NSCLC, although rare false negativity may occur using such samples.