肝细胞癌患者血清外泌体microRNA表达鉴定  

Exosomal MicroRNA Expression Identification in Hepatocellular Carcinoma Patients Serum

作　　者：纪奇峰[1,2] 祝春来 王伟[2,3] 毛壮 汪钦 成珊[1,2] 颜真 JI Qi-feng;ZHU Chun-lai;WANG Wei;MAO Zhuang;WANG Qin;CHENG Shan;YAN Zhen(Department of Pharmacogenomics,School of Pharmacy,Fourth Military Medical University,Xi'an,Shaanxi,710032,China;State Key Laboratory of Cancer Biology,Xi'an,Shaanxi,710032,China;Department of Pharmacy administration and Drug Information,School of Pharmacy,Fourth Military Medical University,Xi'an,Shaanxi,710032,China;Cadet Brigade,Fourth MilitaryMedical University,Xi'an,Shaanxi,710032,China)

机构地区：[1]第四军医大学药学院药物基因组学教研室,陕西西安710032 [2]肿瘤生物学国家重点实验室,陕西西安710032 [3]第四军医大学药学院药事管理与药物信息学教研室,陕西西安710032 [4]第四军医大学学员一旅一营三连,陕西西安710032

出　　处：《现代生物医学进展》2018年第22期4213-4218,共6页Progress in Modern Biomedicine

基　　金：肿瘤生物学国家重点实验室自由探索项目(CBSKL2015Z14);陕西省科技统筹创新工程项目(2016KTCL03-09)

摘　　要：目的:探讨肝癌细胞外泌体中差异表达的microRNAs(miRNAs)在肝细胞癌(HCC)诊断中的应用价值。方法:通过高通量测序筛选肝癌细胞外泌体中差异表达的miRNAs。实时定量PCR验证差异表达分子;检测差异表达的miRNAs在健康人(Health)、慢性乙型肝炎患者(CHB)、肝硬化患者(LC)及乙型肝炎病毒阳性的肝细胞癌患者(HCC)血清外泌体中的表达。结果:高通量测序筛选到肝癌细胞外泌体中差异表达的miRNA共88种,其中58种表达上调,30种表达下调。选择其中8种差异表达的miRNAs进行q RT-PCR验证,结果显示,此8种miRNAs在细胞上清外泌体、细胞内、癌与癌旁组织中的表达趋势与测序结果一致。miR-221-3p和miR-224-5p在HCC组外泌体中的表达水平显著高于Health组、CHB组和LC组(P<0.01),miR-124-3p和let-7a-5p在HCC组外泌体中的表达水平显著低于其他各组(P<0.05)。四个组中,miR-21-5p、miR-191-5p、miR-34a-5p和miR-122-5p的表达水平不存在显著性差异(P>0.05)。结论:血清外泌体中的miR-221-3p、miR-224-5p、miR-124-3p和let-7a-5p可能成为肝细胞癌的候选标志物。Objective: To explore the possible clinical application of differentially expressed exosomal miRNAs in hepatocellular carcinoma (HCC). Methods: The differential expressed miRNAs from HCC cells exosomes were screened by Next Generation Sequencing. The expression levels of differentially expressed miRNAs were confirmed by quantitative real-time PCR and evaluated in healthy controls (Health), hepatitis B patients (CHB), liver cirrhosis patients (LC) and hepatitis B positive HCC patients. Results: Eighty-eight differentially expressed miRNAs were identified in exosomes of HCC cell lines through highthroughput sequencing, including 58 up-regulated and 30 down-regulated miRNAs. Eight differentially expressed miRNAs were verified by qRT-PCR. It showed that miRNA expression was consistent with the sequencing results in either cell line exosomes or in cell lines, also in tumor tissues vs. adjacent tissues. The expression levels of miR-221-3p and miR-224-5p in exosomes were significantly higher in HCC patients than those in Health, CHB or LC group(P<0.01). Furthermore, the expression levels of exosomal miR-124-3p and let-7a-5p were lower in HCC than Health, CHB or LC group (P<0.05). There was no significant difference in the expression of miR-21-5p, miR-191-5p, miR-34a-5p and miR-122-5p among the four groups (P>0.05). Conclusion: Serum exosomal miR-221-3p, miR-224-5p, miR-124-3p and let-7a-5p have the possibility to become canditate biomarkers for hepatocellular carcinoma diagnosis.

关 键 词：肝细胞癌 外泌体 MIRNAS 肿瘤标志物 

分 类 号：R-33[医药卫生] R735.7