摘要
目的筛选肺癌骨转移患者血清外泌体中差异表达的mRNA,分析其作为肺癌骨转移标志物的诊断价值。方法回顾性研究2016-01-01-2019-07-31上海交通大学附属第六人民医院肺癌骨转移患者临床资料,挑选3例肺癌患者发生骨转移前及骨转移后共6例自身对照样本进行高通量测序,采用基因本体论(GO)及京都基因和基因组百科全书(KEGG)对差异基因进行生物信息学分析。通过qPCR对同期本院168例肺癌患者血清外泌体样本中差异最显著的10个基因进行验证。采用受试者工作特征曲线(ROC)评估外泌体差异基因相对表达量对肺癌骨转移的预测价值。结果高通量测序结果发现461条差异表达基因。GO及KEGG分析结果显示,他们参与骨骼肌细胞分化、端粒酶活性的调节、TGF-β信号通路和丙戊酸盐代谢等多种骨转移相关的生物学进程。实时荧光定量PCR(qRT-PCR)验证结果发现,核因子-κB抑制因子α(NFKBIA)和亮氨酸拉链EF-hand结构域跨膜蛋白1(LETMD1)基因在肺癌骨转移组中表达上调,P<0.05。ROC分析结果显示,NFKBIA和LETMD1曲线下面积(AUC)分别为0.788和0.708。进一步将2个差异基因与肺癌血清学诊断标志物CA125、Cyfra211联合分析,发现AUC进一步提高至0.822。结论肺癌骨转移患者血清外泌体中NFKBIA和LETMD1显著高表达,检测血清外泌体NFKBIA和LETMD1表达水平对诊断肺癌骨转移有一定参考价值,可作为肺癌骨转移患者诊断的潜在参考指标。
Objective To screen out the differentially expressed mRNA in serum exosomes of lung cancer patients with bone metastasis and identify its diagnostic value as a new diagnostic biomarker.Methods The clinical data of lung cancer patients with bone metastasis from January 1,2016 to July 31,2019 in the Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University were retrospectively studied.A total of 6 samples from 3 lung cancer patients before and after bone metastasis were selected for high-throughput sequencing.The bioinformatics analysis was performed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.Quantitative real-time PCR(qPCR)were used to identify 10 genes with the most significant differences in 168 clinical serum exosomes.ROC curve was used to evaluate the predictive value of the relative expression of exosome-derived differential mRNAs for the lung cancer patients with bone metastasis.Results A total of 461 differentially expressed genes were identified by high-throughput sequencing.The results of GO and KEGG analyses showed that they were involved in skeletal muscle cell differentiation,regulation of telomerase activity,TGF-beta signaling pathway,propanoate metabolism and other biological processes related to bone metastasis.The results of qPCR showed that NFkB inhibitor alph(NFKBIA)and LETMD1 domain containing 1(LETMD1)were up-regulated in lung cancer patients with bone metastasis(P<0.05).Receiver operating characteristic curve(ROC)showed that the area under curves(AUC)of NFKBIA and LETMD1 were 0.788 and 0.708,respectively.Furthermore,the AUC was further increased to 0.822 when NFKBIA and LETMD1 were combined with CA125 andCyfra211.Conclusion The mRNA expression levels of NFKBIA and LETMD1 in serum exosomes of lung cancer patients with bone metastasis are significantly up-regulated,which can be used as a potential biomarker for the diagnosis of bone metastasis of lung cancer.
作者
夏前林
付亚文
滕小艳
魏丽荣
杜玉珍
XIA Qian-lin;FU Ya-wen;TENG Xiao-yan;WEI Li-rong;DU Yu-zhen(Department of Laboratory Medicine,Shanghai Jiao Tong University Affiliated Sixth People's Hospital,Shanghai 201306,China)
出处
《中华肿瘤防治杂志》
CAS
北大核心
2021年第18期1392-1397,共6页
Chinese Journal of Cancer Prevention and Treatment
关键词
肺癌
骨转移
外泌体
肿瘤标志物
lung cancer
bone metastasis
exosomes
tumor markers
作者简介
第一作者:夏前林,男,上海人,硕士,初级检验技师,主要从事肿瘤生化与分子诊断的研究工作。E-mail:xiaqianlin@foxmail.com;通信作者:杜玉珍,女,上海人,博士,主任技师,主要从事肿瘤生化与分子诊断的研究工作E-mail:duyuzhen2005@163.com
