肺癌患者血浆外泌体microRNAs标志物的筛选

doi:10.3969/j.issn.1004-616x.2022.06.001

摘要/Abstract

摘要： 目的：筛选肺癌患者血浆外泌体中的异常microRNA(miRNA)，以期为肺癌筛查及辅助诊断提供新的标志物和方向。方法：选择6名肺癌患者(腺癌3例、鳞癌2例，小细胞肺癌1例)和4名健康对照者血浆样本，采用差速离心法提取血浆外泌体，采用Agilent Bioanalyzer对抽提的RNA进行质量检验，对血浆外泌体miRNA进行高通量测序分析，采用TargetScan、PITA和miRNAorg数据库进行靶基因预测，应用R软件进行靶基因的KEGG和GO分析。结果：miRNA表达分析发现，与正常对照者比较，肺癌患者血浆外泌体中有142种miRNAs表达异常，其中62种miRNAs表达上调，80种miRNAs表达下调。GO分析显示，这些miRNAs的靶基因主要参与了以DNA为模板的转录、转录调控及信号转导等。KEGG分析显示，这些miRNAs的靶基因主要参与轴突导向信号通路、钙信号通路、肌动蛋白细胞骨架调节等。结论：通过对肺癌患者和健康对照者血浆外泌体miRNAs进行高通量测序，初步筛选出差异表达miRNA，经生物信息学分析，推测其对肺癌转移具有良好预测潜力，有望成为肺癌筛查和辅助诊断的候选生物标志物。

关键词: 肺癌, 筛查, 外泌体, 肿瘤标志物, 微小RNA

Abstract: OBJECTIVE:Existence of miRNAs in plasma exosomes of lung cancer patients were screened to develop new methods for the screening and auxiliary diagnosis of lung cancers. METHODS:Plasma samples were collected from 6 lung cancer patients (3 adenocarcinomas,2 squamous carcinomas and 1 small cell lung cancer) and 4 healthy controls. Agilent Bioanalyzer was used for qualitative inspection of the extracted RNA. Plasma exosomal miRNA was analyzed by high-throughput sequencing. Target gene prediction was performed by TargetScan,PITA and miRNAorg database,and R software was applied for KEGG and GO analysis of target genes. RESULTS:The exosomal differential miRNA expression analyses showed that 142 miRNAs were expressed abnormally in the lung cancer group compared with to the normal group,with 62 miRNAs up-regulated and 80 miRNAs down-regulated. GO analysis showed that these abnormally-expressed (target) genes were mainly involved in DNA-based template transcription,transcriptional regulation and signal transduction,etc. The target KEGG analysis indicated that these target genes were mainly involved in axon guidance signaling pathway,calcium signaling pathway,and actin cytoskeleton regulation. CONCLUSION:Our data suggest that differentially expressed miRNAs may have good predictive potential for lung cancer metastasis. In addition,they may be candidate biomarkers for the screening and diagnosis of lung cancers.

Key words: lung cancer, screening, exosome, tumor marker, microRNA

中图分类号:

R114

刘霞, 高孜博, 丁丽华, 吴拥军. 肺癌患者血浆外泌体microRNAs标志物的筛选[J]. 癌变·畸变·突变, 2022, 34(6): 407-412.

LIU Xia, GAO Zibo, DING Lihua, WU Yongjun. Screening of plasma exosome microRNAs in patients with lung cancer[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2022, 34(6): 407-412.

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