Comprehensive analysis of NGS and ARMS-PCR for detecting EGFR mutations based on 4467 cases of NSCLC patients

Journal of Cancer Research and Clinical Oncology - Tập 148 - Trang 321-330 - 2021
Changlong He1, Chengcheng Wei2,3, Jun Wen2, Shi Chen2, Ling Chen1, Yue Wu4, Yifan Shen2, Huili Bai2, Yangli Zhang2, Xueping Chen2, Xiaosong Li2
1Key Laboratory of Molecular Biology for Infectious Diseases (Ministry of Education), Department of Infectious Diseases, Institute for Viral Hepatitis, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China
2Clinical Molecular Medicine Testing Center, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China
3Department of Urology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
4Oncology Department, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China

Tóm tắt

By comparing the detection rate and type of targeted gene mutations in non-small cell lung cancer (NSCLC) between amplification refractory mutation system PCR (ARMS-PCR) and next-generation sequencing (NGS), the characteristics and application advantages of non-small cell lung cancer detection are explained, providing a basis for clinicians to effectively select the corresponding detection methods. The cases of targeted genes for lung cancer were selected from the First Affiliated Hospital of Chongqing Medical University from January 2016 to October 2020. A sample of 4467 cases was selected, and they were diagnosed with NSCLC by Pathological biopsy. Sample sources include surgical resection, bronchoscope biopsy, metastatic biopsy, blood, sputum, cytology of pleural effusion. Among them, 3665 cases were detected by ARMS-PCR technique, and 802 cases were detected by NGS technology. The detection rate and type of ARMS-PCR and NGS techniques for EGFR gene mutations (including exon 18, exon 19, exon 20, exon 21 and so on) in different NSCLC samples were compared, respectively. The total mutation rate of EGFR gene detected by ARMS-PCR was 47.6% while 42.4% detected by NGS which indicated that there was a significant difference between the two methods in detecting total mutation of EGFR gene (P < 0.001). In different exons, the EGFR mutation rate detected by two methods is various. The mutation rate of exon 19 by ARMS-PCR detection was evidently higher than that of NGS detection, while the mutation rate of exons 20 and 21 by ARMS-PCR detection were statistically significantly lower than that of NGS detection. Moreover, the multiple mutation rate detected by NGS was 16.3% which was much higher than the 2.7% detected by ARMS-PCR with statistically different. It showed that NGS could direct the drug use for the resistant patients. However, some rare loci could be detected by NGS but the importance and directed meaning are still unknown and the number of rare mutations is rare too. Further research on new biomarkers and technique is still needed for early diagnosis, directing drug use and assessing the therapy prognosis.

Tài liệu tham khảo

Bray F et al (2018) Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 68: 394–424. https://doi.org/10.3322/caac.21492 Chen M, Zhao H (2019) Next-generation sequencing in liquid biopsy: cancer screening and early detection. Hum Genomics 13:34. https://doi.org/10.1186/s40246-019-0220-8 Chen W, et al (2016) Cancer statistics in China, 2015. CA Cancer J Clin. 66: 115–132. https://doi.org/10.3322/caac.21338 Coco S et al (2015) Next generation sequencing in non-small cell lung cancer: new avenues toward the personalized medicine. Curr Drug Targets 16:47–59. https://doi.org/10.2174/1389450116666141210094640 Crowley E, Di Nicolantonio F, Loupakis F, Bardelli A (2013) Liquid biopsy: monitoring cancer-genetics in the blood. Nat Rev Clin Oncol 10:472–484. https://doi.org/10.1038/nrclinonc.2013.110 Dahabreh IJ et al (2010) Somatic EGFR mutation and gene copy gain as predictive biomarkers for response to tyrosine kinase inhibitors in non-small cell lung cancer. Clin Cancer Res 16:291–303. https://doi.org/10.1158/1078-0432.Ccr-09-1660 Dajac J, Kamdar J, Moats A, Nguyen B (2016) To screen or not to screen: low dose computed tomography in comparison to chest radiography or usual care in reducing morbidity and mortality from lung cancer. Cureus 8:e589. https://doi.org/10.7759/cureus.589 Ferlay J et al (2015) Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer 136:E359-386. https://doi.org/10.1002/ijc.29210 Gazzeri S (2018) Nuclear EGFR: a new mode of oncogenic signalling in cancer. Biol Aujourdhui 212:27–33. https://doi.org/10.1051/jbio/2018016 Gerlinger M et al (2012) Intratumor heterogeneity and branched evolution revealed by multiregion sequencing. N Engl J Med 366:883–892. https://doi.org/10.1056/NEJMoa1113205 Graham RP et al (2018) Worldwide frequency of commonly detected EGFR mutations. Arch Pathol Lab Med 142:163–167. https://doi.org/10.5858/arpa.2016-0579-CP Hassanein M et al (2012) The state of molecular biomarkers for the early detection of lung cancer. Cancer Prev Res 5:992–1006. https://doi.org/10.1158/1940-6207.Capr-11-0441 Kimura H et al (2006) Detection of epidermal growth factor receptor mutations in serum as a predictor of the response to gefitinib in patients with non-small-cell lung cancer. Clin Cancer Res 12:3915–3921. https://doi.org/10.1158/1078-0432.Ccr-05-2324 Kruglyak KM, Lin E, Ong FS (2016) Next-generation sequencing and applications to the diagnosis and treatment of lung cancer. Adv Exp Med Biol 890:123–136. https://doi.org/10.1007/978-3-319-24932-2_7 Lee SH et al (2015) Analysis of mutations in epidermal growth factor receptor gene in Korean patients with non-small cell lung cancer: summary of a nationwide survey. J Pathol Transl Med 49:481–488. https://doi.org/10.4132/jptm.2015.09.14 Liam CK, Wahid MI, Rajadurai P, Cheah YK, Ng TS (2013) Epidermal growth factor receptor mutations in lung adenocarcinoma in Malaysian patients. J Thorac Oncol 8:766–772. https://doi.org/10.1097/JTO.0b013e31828b5228 Liang C et al (2018) Detection of rare mutations in EGFR-ARMS-PCR-negative lung adenocarcinoma by Sanger sequencing. Yonsei Med J 59:13–19. https://doi.org/10.3349/ymj.2018.59.1.13 Liu J, Zhao R, Zhang J, Zhang J (2015) ARMS for EGFR mutation analysis of cytologic and corresponding lung adenocarcinoma histologic specimens. J Cancer Res Clin Oncol 141:221–227. https://doi.org/10.1007/s00432-014-1807-z Ma Y et al (2020) Oncogenic genetic alterations in non-small-cell lung cancer (NSCLC) in Southwestern China. Cancer Manag Res 12:10861–10874. https://doi.org/10.2147/cmar.S266069 Masago K et al (2015) Next-generation sequencing of tyrosine kinase inhibitor-resistant non-small-cell lung cancers in patients harboring epidermal growth factor-activating mutations. BMC Cancer 15:908. https://doi.org/10.1186/s12885-015-1925-2 Morganti S et al (2019) Next Generation Sequencing (NGS): a revolutionary technology in pharmacogenomics and personalized medicine in cancer. Adv Exp Med Biol 1168:9–30. https://doi.org/10.1007/978-3-030-24100-1_2 Nakata A et al (2015) Elevated β-catenin pathway as a novel target for patients with resistance to EGF receptor targeting drugs. Sci Rep 5:13076. https://doi.org/10.1038/srep13076 Pantel K, Alix-Panabières C (2013) Real-time liquid biopsy in cancer patients: fact or fiction? Can Res 73:6384–6388. https://doi.org/10.1158/0008-5472.Can-13-2030 Penzel R et al (2011) EGFR mutation detection in NSCLC–assessment of diagnostic application and recommendations of the German panel for mutation testing in NSCLC. Virchows Arch 458:95–98. https://doi.org/10.1007/s00428-010-1000-y Rangachari D et al (2015) Experience with targeted next generation sequencing for the care of lung cancer: insights into promises and limitations of genomic oncology in day-to-day practice. Cancer Treat Commun 4:174–181. https://doi.org/10.1016/j.ctrc.2015.10.004 Roskoski R Jr (2014) ErbB/HER protein-tyrosine kinases: Structures and small molecule inhibitors. Pharmacol Res 87:42–59. https://doi.org/10.1016/j.phrs.2014.06.001 Shao D et al (2016) A targeted next-generation sequencing method for identifying clinically relevant mutation profiles in lung adenocarcinoma. Sci Rep 6:22338. https://doi.org/10.1038/srep22338 Shi Y et al (2014) A prospective, molecular epidemiology study of EGFR mutations in Asian patients with advanced non-small-cell lung cancer of adenocarcinoma histology (PIONEER). J Thorac Oncol 9:154–162. https://doi.org/10.1097/jto.0000000000000033 Shi Y et al (2015) Molecular epidemiology of EGFR mutations in Asian patients with advanced non-small-cell lung cancer of adenocarcinoma Histology-Mainland China subset analysis of the PIONEER study. PLoS One 10:e0143515. https://doi.org/10.1371/journal.pone.0143515 Sholl LM et al (2010) EGFR mutation is a better predictor of response to tyrosine kinase inhibitors in non-small cell lung carcinoma than FISH, CISH, and immunohistochemistry. Am J Clin Pathol 133:922–934. https://doi.org/10.1309/ajcpst1cthzs3psz Tan AC et al (2020) Utility of incorporating next-generation sequencing (NGS) in an Asian non-small cell lung cancer (NSCLC) population: Incremental yield of actionable alterations and cost-effectiveness analysis. Lung Cancer 139:207–215. https://doi.org/10.1016/j.lungcan.2019.11.022 Thomas A, Rajan A, Lopez-Chavez A, Wang Y, Giaccone G (2013) From targets to targeted therapies and molecular profiling in non-small cell lung carcinoma. Ann Oncol 24:577–585. https://doi.org/10.1093/annonc/mds478 Travis WD et al (2015) The 2015 World Health Organization classification of lung tumors: impact of genetic, clinical and radiologic advances since the 2004 classification. J Thorac Oncol 10:1243–1260. https://doi.org/10.1097/jto.0000000000000630 Won JK et al (2015) Concomitant ALK translocation and EGFR mutation in lung cancer: a comparison of direct sequencing and sensitive assays and the impact on responsiveness to tyrosine kinase inhibitor. Ann Oncol 26:348–354. https://doi.org/10.1093/annonc/mdu530 Xuan J, Yu Y, Qing T, Guo L, Shi L (2013) Next-generation sequencing in the clinic: promises and challenges. Cancer Lett 340:284–295. https://doi.org/10.1016/j.canlet.2012.11.025 Yohe S, Thyagarajan B (2017) Review of clinical next-generation sequencing. Arch Pathol Lab Med 141:1544–1557. https://doi.org/10.5858/arpa.2016-0501-RA Zhou J et al (2016) Prevalence and clinical profile of EGFR mutation in non-small-cell lung carcinoma patients in Southwest China. Asian Pac J Cancer Prev 17:965–971. https://doi.org/10.7314/apjcp.2016.17.3.965 Zhou YC et al (2020) Analysis of EGFR mutation and clinical features of lung cancer in Yunnan. Zhonghua zhong liu za zhi [Chinese journal of oncology] 42:729–734. https://doi.org/10.3760/cma.j.cn112152-20200313-00201