非小细胞肺癌组织EGFR/ALK/ROS1基因联合检测的临床意义
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摘要
目的探讨非小细胞肺癌(NSCLC)患者表皮生长因子受体(EGFR),间变性淋巴瘤激酶(ALK)和肉瘤致癌因子-受体酪氨酸激酶(ROS1)三种基因突变联合检测的临床意义。方法收集2017年1月～2018年7月患者NSCLC组织标本146例,采用过柱法提取DNA和RNA,突变检测采用RT-PCR法和扩增阻滞突变系统(ARMS)法。结果EGFR/ALK/ROS1三联检的突变频率为41.8%(61/146),明显高于单基因检测突变率:EGFR (33.6%,49/146),ALK(6.8%,10/146)和ROS1(2.7%,4/146)。EGFR的不同突变类型的检出率如下:19Del 52.9%(27/51),L858R41.2%(21/51),G719X,L861Q5.9%(3/51);其中还检出四例双突变标本:19Del+L858R(1例),19Del+G719X,L861Q(1例),ALK+19Del(1例),ROS1+L858R(1例)。EGFR/ALK/ROS1阳性标本中女性(56.7%,30/53)明显高于男性(33.3%,31/93),两者比较差异有统计学意义(χ~2=7.516,P=0.006),肺腺癌(47.9%,58/121)明显高于鳞癌(4.8%,1/21),差异具有统计学意义(χ~2=13.733,P=0.000)。不同来源组织突变检出率为:手术切除肿瘤组织标本41.0%(32/78),细针穿刺活检及支气管镜刷检细胞学等标本43.4%(23/53),胸腔积液标本40.0%(6/15),三者比较差异无统计学意义(P>0.05)。结论在NSCLC基因突变中:EGFR/ALK/ROS1三联检的突变频率明显高于EGFR,ALK和ROS1单基因检测。EGFR常见突变基因为19Ex Del和21Ex L858R。女性突变率明显高于男性。EGFR/ALK/ROS1的联合检测可一次获得更多基因突变信息,为靶向用药提供更多选择,尤其是对于临床稀有标本意义更大。
Objective To explore the clinical significance of combined detection of three gene mutations in non-small cell lung cancer(NSCLC)patients:epidermal growth factor receptor(EGFR),mesenchymal lymphoma kinase(ALK)and sarcomao ncogene-receptor tyrosine kinase(ROS1).Methods From January 2017 to July 2018,146 patients with NSCLC tissue were collected in Shaanxi Province People's Hospital.DNA and RNA were extracted by centrifugal mini column methods.RT-PCR and amplification block mutation system(ARMS)method were employed in the mutation detection.Results The gene mutation rate of EGFR/ALK/ROS1(41.8%,61/146)was significantly higher than that of EGFR(33.6%,49/146),ALK(6.8%,10/146)and ROS1(2.7%,4/146).The mutation rate of different mutation types of EGFR gene were as follows:deletions at exon 19 was 52.9%(27/51),21 Ex L858 Rwas 41.2%(21/51),18 Ex G719 X,21 Ex L861 Qwas 5.9%(3/51);Also co-mutation of 19 Del+ L858 R(1 sample),19 Del+ G719 X,L861 Q(1 sample),ALK+19 Del(1 sample),and ROS1+L858 R(1 sample)in EGFR were detected.Among EGFR/ALK/ROS1 positive samples,women(56.7%,30/53)were higher than men(33.3%,31/93),the comparison was statistically significant(χ~2=7.516,P=0.006),and lung adenocarcinoma(47.9%,58/121)was significantly higher than squamous cancer(4.8%,1/21),the comparison was statistically significant(χ~2=13.733,P=0.000).The mutation detection rate of different source organizations were as follows:large tissue specimens(41.0%,32/78),fine needle biopsy and bronchoscopic brush cytology(43.4%,23/53),and pleural water specimens(40.0%,6/15).Conclusion In the NSCLC gene mutation,the mutation frequency of EGFR/ALK/ROS1 was significantly higher than that of EGFR,ALK and ROS1 gene detection,and the common mutation genes of EGFR were 19 Ex Del and 21 Ex L858 R.Female mutation rate was significantly higher than male.The combined detection of EGFR/ALK/ROS1 can obtain more information of gene mutation at one time,providing more choices for targeted drugs,especially for rare clinical specimens.
Objective To explore the clinical significance of combined detection of three gene mutations in non-small cell lung cancer(NSCLC)patients:epidermal growth factor receptor(EGFR),mesenchymal lymphoma kinase(ALK)and sarcomao ncogene-receptor tyrosine kinase(ROS1).Methods From January 2017 to July 2018,146 patients with NSCLC tissue were collected in Shaanxi Province People's Hospital.DNA and RNA were extracted by centrifugal mini column methods.RT-PCR and amplification block mutation system(ARMS)method were employed in the mutation detection.Results The gene mutation rate of EGFR/ALK/ROS1(41.8%,61/146)was significantly higher than that of EGFR(33.6%,49/146),ALK(6.8%,10/146)and ROS1(2.7%,4/146).The mutation rate of different mutation types of EGFR gene were as follows:deletions at exon 19 was 52.9%(27/51),21 Ex L858 Rwas 41.2%(21/51),18 Ex G719 X,21 Ex L861 Qwas 5.9%(3/51);Also co-mutation of 19 Del+ L858 R(1 sample),19 Del+ G719 X,L861 Q(1 sample),ALK+19 Del(1 sample),and ROS1+L858 R(1 sample)in EGFR were detected.Among EGFR/ALK/ROS1 positive samples,women(56.7%,30/53)were higher than men(33.3%,31/93),the comparison was statistically significant(χ~2=7.516,P=0.006),and lung adenocarcinoma(47.9%,58/121)was significantly higher than squamous cancer(4.8%,1/21),the comparison was statistically significant(χ~2=13.733,P=0.000).The mutation detection rate of different source organizations were as follows:large tissue specimens(41.0%,32/78),fine needle biopsy and bronchoscopic brush cytology(43.4%,23/53),and pleural water specimens(40.0%,6/15).Conclusion In the NSCLC gene mutation,the mutation frequency of EGFR/ALK/ROS1 was significantly higher than that of EGFR,ALK and ROS1 gene detection,and the common mutation genes of EGFR were 19 Ex Del and 21 Ex L858 R.Female mutation rate was significantly higher than male.The combined detection of EGFR/ALK/ROS1 can obtain more information of gene mutation at one time,providing more choices for targeted drugs,especially for rare clinical specimens.
引文
[1] Torre LA,Bray F,Siegel RL,et al.Global cancer statistics,2012[J].CA-A Cancer Journal Clinicians,2015,65(2):87-108.
[2]叶肖燕.非小细胞肺癌患者表皮生长因子受体基因突变特征及意义[J].中华实用诊断及治疗杂志,2016,30(1):49-51.Ye XY.Characteristics and significance of epidermal growth factor receptor gene mutation in non-small cell lung cancer[J].J Chin Practical Diagnosis and Therapy,2016,30(1):49-51.
[3] She J,Yang P,Hong QY,et al.Lung cancer in China challenges and interventions[J].Chest,2013,143(4):1117-1126.
[4]赵建国,熊建萍.非小细胞肺癌驱动基因研究进展[J].中国肺癌杂志,2015,18(1):42-47.Zhao JG,Xiong JP.Advances on driver oncogenes of non-small cell lung cancer[J].Chin J Lung Cancer,2015,18(1):42-47.
[5]黎谢梦丹,罗凯,吴顺芳,等.华南地区非小细胞肺癌患者肿瘤组织EGFR,ALK和ROS1基因突变分析[J].现代检验医学杂志,2017,32(5):16-18,23.Lixie MD,Luo K,Wu SF,et al.Mutation analysis of EGFR,ALK and ROS1in tumor tissues of patients with non-small cell lung cancer in south of China[J].J Mod Lab Med,2017,32(5):16-18,23.
[6] Yamamoto H,Toyooka S,Mitsudomi T.Impact of EGFR mutation analysis in non-small cell lung cancer[J].Lung Cancer,2018,63(3):315-324.
[7] Shaw AT,Yeap BY,Solomon BJ,et al.Effect of crizotinib on overall survival in patients with advanced non-small-cell lung cancer harbouring ALK gene rearrangement:a retrospective analysis[J].Lancet Oncol,2011,12(11):1004-1012.
[8] Ou SH,Tan J,Yen Y,et al.ROS1as a“druggable”receptor tyrosine kinase:lessons learned from inhibiting the ALK pathway[J].Expert Rev Anticancer Ther,2012,12(4):447-456.
[9] Sholl L.Molecular diagnostics of lung cancer in the clinic[J].Transl Lung Cancer Res,2017,6(5):560-569.
[10] Shao AT,Ou SH,Bang YJ,et al.Crizotinib in ROS1rearranged non-small cell lung cancer[J].N Engl J Med,2014,371(21):1963-1971.
[11]中国非小细胞肺癌患者表皮生长因子受体基因突变检测专家组.中国非小细胞肺癌患者表皮生长因子受体基因突变检测专家共识(2016版)[J].中华病理学杂志,2016,45(4):217-220.Expert Group for Detection of Epidermal Growth Fa-ctor Receptor Gene Mutation in Non-small Cell Lung Cancer Patients in China.Expert consensus on the detection of epidermal growth factor receptor gene mutation in non-small cell lung cancer patients in China(2016edition)[J].Chin J Pathol,2016,45(4):217-220.
[12] Campbell JD,Alexandrov A,Kim J,et al.Distinct pa-tterns of somatic genome alterations in lung adenocarcinomas and squamous cell carcinomas[J].Nat Genet,2016,48(6):607-616.
[13] Li SY,Choi YL,Gong ZL,et al.Comprehensive characterization of oncogenic drivers in Asian lung adenocarcinoma[J].J Thorac Oncol,2016,11(12):2129-2140.
[14]杨长绍,杨延龙,丁晓洁,等.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[J].临床与实验病理学杂志,2017,33(11):1183-1187.Yang CS,Yang YL,Ding XJ,et al.Driver gene mutations among non-small cell lung cancer tissues and its clinical significance[J].J Clin Exp Pathol,2017,33(11):1183-1187.
[15] Doss GP,Rajith B,Chakraborty C,et al.Structural signature of the G719S-T790M double mutation in the EGFR kinase domain and its response to inhibitors[J].Sci Rep,2014,4:5868.
[16] Chiu CH,Yang CT,Shih JY,et al.Epidermal growth factor receptor tyrosine kinase inhibitor treatment response in advanced lung adenocarcinomas with G719X/L861Q/S768Imutations[J].J Thorac Oncol,2015,10(5):793-799.
[17] Barnet MB,O'Toole S,Horvath LG,et al.EGFR-CoMutated advanced NSCLC and response to EGFR tyrosine kinase inhibitors[J].J Thorac Oncol,2017,12(3):585-590.
[2]叶肖燕.非小细胞肺癌患者表皮生长因子受体基因突变特征及意义[J].中华实用诊断及治疗杂志,2016,30(1):49-51.Ye XY.Characteristics and significance of epidermal growth factor receptor gene mutation in non-small cell lung cancer[J].J Chin Practical Diagnosis and Therapy,2016,30(1):49-51.
[3] She J,Yang P,Hong QY,et al.Lung cancer in China challenges and interventions[J].Chest,2013,143(4):1117-1126.
[4]赵建国,熊建萍.非小细胞肺癌驱动基因研究进展[J].中国肺癌杂志,2015,18(1):42-47.Zhao JG,Xiong JP.Advances on driver oncogenes of non-small cell lung cancer[J].Chin J Lung Cancer,2015,18(1):42-47.
[5]黎谢梦丹,罗凯,吴顺芳,等.华南地区非小细胞肺癌患者肿瘤组织EGFR,ALK和ROS1基因突变分析[J].现代检验医学杂志,2017,32(5):16-18,23.Lixie MD,Luo K,Wu SF,et al.Mutation analysis of EGFR,ALK and ROS1in tumor tissues of patients with non-small cell lung cancer in south of China[J].J Mod Lab Med,2017,32(5):16-18,23.
[6] Yamamoto H,Toyooka S,Mitsudomi T.Impact of EGFR mutation analysis in non-small cell lung cancer[J].Lung Cancer,2018,63(3):315-324.
[7] Shaw AT,Yeap BY,Solomon BJ,et al.Effect of crizotinib on overall survival in patients with advanced non-small-cell lung cancer harbouring ALK gene rearrangement:a retrospective analysis[J].Lancet Oncol,2011,12(11):1004-1012.
[8] Ou SH,Tan J,Yen Y,et al.ROS1as a“druggable”receptor tyrosine kinase:lessons learned from inhibiting the ALK pathway[J].Expert Rev Anticancer Ther,2012,12(4):447-456.
[9] Sholl L.Molecular diagnostics of lung cancer in the clinic[J].Transl Lung Cancer Res,2017,6(5):560-569.
[10] Shao AT,Ou SH,Bang YJ,et al.Crizotinib in ROS1rearranged non-small cell lung cancer[J].N Engl J Med,2014,371(21):1963-1971.
[11]中国非小细胞肺癌患者表皮生长因子受体基因突变检测专家组.中国非小细胞肺癌患者表皮生长因子受体基因突变检测专家共识(2016版)[J].中华病理学杂志,2016,45(4):217-220.Expert Group for Detection of Epidermal Growth Fa-ctor Receptor Gene Mutation in Non-small Cell Lung Cancer Patients in China.Expert consensus on the detection of epidermal growth factor receptor gene mutation in non-small cell lung cancer patients in China(2016edition)[J].Chin J Pathol,2016,45(4):217-220.
[12] Campbell JD,Alexandrov A,Kim J,et al.Distinct pa-tterns of somatic genome alterations in lung adenocarcinomas and squamous cell carcinomas[J].Nat Genet,2016,48(6):607-616.
[13] Li SY,Choi YL,Gong ZL,et al.Comprehensive characterization of oncogenic drivers in Asian lung adenocarcinoma[J].J Thorac Oncol,2016,11(12):2129-2140.
[14]杨长绍,杨延龙,丁晓洁,等.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[J].临床与实验病理学杂志,2017,33(11):1183-1187.Yang CS,Yang YL,Ding XJ,et al.Driver gene mutations among non-small cell lung cancer tissues and its clinical significance[J].J Clin Exp Pathol,2017,33(11):1183-1187.
[15] Doss GP,Rajith B,Chakraborty C,et al.Structural signature of the G719S-T790M double mutation in the EGFR kinase domain and its response to inhibitors[J].Sci Rep,2014,4:5868.
[16] Chiu CH,Yang CT,Shih JY,et al.Epidermal growth factor receptor tyrosine kinase inhibitor treatment response in advanced lung adenocarcinomas with G719X/L861Q/S768Imutations[J].J Thorac Oncol,2015,10(5):793-799.
[17] Barnet MB,O'Toole S,Horvath LG,et al.EGFR-CoMutated advanced NSCLC and response to EGFR tyrosine kinase inhibitors[J].J Thorac Oncol,2017,12(3):585-590.