高通量测序平台研究非小细胞肺癌热点基因突变情况分析  

High-throughput Sequencing Platform to Study Hot Spot Gene Mutations in Non-small Cell Lung Cancer

作　　者：于丽丽 YU Lili(Clinical laboratory,Jiamusi Central Hospital,Jiamusi City,Heilongjiang Province 154001)

机构地区：[1]黑龙江省佳木斯市中心医院检验科,154001

出　　处：《医学理论与实践》2021年第19期3320-3322,共3页The Journal of Medical Theory and Practice

摘　　要：目的:探讨与分析高通量测序平台研究非小细胞肺癌热点基因突变情况。方法:选择2019年11月-2020年11月在本院肿瘤科诊治的非小细胞肺癌患者110例作为肺癌组,同期选择健康体检者110例作为对照组,提取血浆组织,采用高通量测序平台分析基因表达差异情况,重点检测热点基因突变情况。结果:两组所有入选者DNA提取体积>40μl,浓度>1ng/μl,总量>55ng,<200nt比例<5%,都符合质控要求,两组对比差异无统计学意义(P>0.05)。两组筛选出显著差异表达的基因共1245个,其中肺癌组表达上调652个,表达下调593个。基因测序与实时荧光定量PCR显示肺癌组的鼠肉瘤病毒致癌基因(Kirsten rat sarcoma viral oncogene,KRAS)、磷脂酰肌醇-3-激酶催化亚基α基因(Phosphoinositide-3-kinase,catalytic,alpha gene,PIK3CA)、肿瘤蛋白P53(Tumor protein P53,TP53)表达水平都高于对照组(P<0.05)。基因测序显示肺癌组的KRAS、PIK3CA、TP53基因突变率分别为30.0%、22.7%、17.3%,高于对照组的3.6%、1.8%和0.9%(P<0.05)。结论:高通量测序平台能有效检出非小细胞肺癌热点基因突变情况,具有很好的应用可行性,值得推广应用。Objective:To explore and analysis the high-throughput sequencing platform to study the hot spot gene mutations in non-small cell lung cancer.Methods:From November 2019 to November 2020,110 cases of patients with non-small cell lung cancer diagnosed and treated in the oncology department of our hospital were selected as the lung cancer group,and 110 cases of healthy people were selected as the control group during the same period.The plasma tissue were extracted and high-throughput sequencing were used.The platform used to analysis the differences in gene expression,focusing on detecting hot-spot gene mutations.Results:The DNA extraction volume of all cases in the two groups were>40μl,the concentration were>1ng/μl,the total amount were>55ng,and the ratio of<200nt were less than 5%,which all met the quality control requirements.There were no statistically significant compared difference between the two groups(P>0.05).A total of 1245 genes with significant differential expression were screened between the two groups,of which 652 genes were up-regulated in the lung cancer group and 593 genes were down-regulated in the lung cancer group.Gene sequencing and real-time fluorescent quantitative PCR showed that the oncogenes of Kirsten rat sarcoma viral oncogene(KRAS),Phosphoinositide-3-kinase,catalytic,alpha gene(PIK3CA),tumor protein P53(TP53)expression levels were higher than those of the control group(P<0.05).Gene sequencing showed that the mutation rates of KRAS,PIK3CA,and TP53 genes in the lung cancer group were 30.0%,22.7%,and 17.3%,respectively,which were higher than 3.6%,1.8%,and 0.9%in the control group(P<0.05).Conclusion:The high-throughput sequencing platform can effectively detect hot-spot gene mutations in non-small cell lung cancer,has good application feasibility,and is worthy of popularization and application.

关 键 词：高通量测序平台 非小细胞肺癌 基因突变 鼠肉瘤病毒致癌基因 磷脂酰肌醇-3-激酶催化亚基α基因 肿瘤蛋白P53 

分 类 号：R734.2[医药卫生—肿瘤] R363[医药卫生—临床医学]