小鼠白血病细胞L1210外泌体微RNA表达谱及其功能分析  

Expression profile and functional analysis of microRNAs in exosomes secreted by murine acute leukemia cells L1210

作　　者：姚烨[1] 黄芳[1] 郝思国[1] 万江波[1] 张文皓[1] 马立元[1] 邓晓辉[1] 陈琳军[1] 

机构地区：[1]上海交通大学医学院附属新华医院血液科,200092

出　　处：《白血病．淋巴瘤》2016年第9期518-525,共8页Journal of Leukemia & Lymphoma

基　　金：国家自然科学基金（81071857）

摘　　要：目的：对白血病细胞外泌体（LCEX）中微 RNA（miRNA）的表达特点及其功能进行分析。方法以小鼠白血病细胞株 L1210为模型，应用密度梯度超速离心法分离其培养上清，获得 LCEX L1210，应用基因芯片技术检测 L1210细胞和 LCEXL1210中 miRNA 的表达，比较二者 miRNA 的表达，对部分差异表达的miRNA 应用实时荧光定量 PCR 进行验证，通过 Gene Ontology 数据库分析。结果在 LCEX L1210中共鉴定出1044种 miRNA，L1210细胞中共鉴定出872种 miRNA，其中732种 miRNA 为两者共有，两者共同表达的 miRNA 占 L1210细胞的83.9%，占 LCEXL1210的70.1%，提示 LCEXL1210中70%以上 miRNA 来自于其母细胞。在 LCEXL1210中有312种 miRNA 在其母体细胞中没有鉴定出，为 LCEXL1210所特有。有些miRNA 在 LCEXL1210中表达明显高于 L1210细胞，如 miR-16-1、miR-210及 miR-195等，提示 LCEX 的miRNA 表达与其母体细胞存在表达差异。对部分表达差异的 miRNA 应用实时荧光定量 PCR 验证显示，这些 LCEX 高表达的 miRNA 参与多种生物学功能及信号转导途径的调控。结论 LCEXL1210所含的miRNA 与其来源细胞 L1210细胞所含的 miRNA 有高度相似性，但有1／3的 miRNA 为其所特有。这些LCEXL1210中高表达的 miRNA 参与多种生物学功能及信号转导途径的调控。Objective To analyze the expression feature and function of microRNAs in exosomes secreted by leukemia cells （LCEX）. Methods The mice leukemia cell line L1210 was taken as the example, and LCEXL1210 was obtained by isolating supernate of L1210 cells through density gradient centrifugation. MicroRNAs isolated from LCEXL1210 were analyzed by microarray analysis, compared with miRNA from L1210 cell line, and then some of miRNAs with different expression were verified by real-time PCR and were analyzed by Gene Ontology （GO） database. Results The number of miRNAs identified in LCEXL1210 was 1 044, and that in L1210 cell line was 872. The number of shared miRNAs between LCEXL1210 and L1210 cell line was 732, accounting for 70.1 % of LCEXL1210 and 83.9 % of L1210 cell line, respectively, which indicated that 70 % of LCEXL1210 was derived from the parental cells. Interestingly, 312 miRNAs in LCEXL1210 were found to be underrepresented in the parental cells, indicating their specificity in LCEXL1210. Some miRNAs were significantly highly expressed in LCEXL1210 compared with those in L1210 cell line, including miR-16-1, miR-210, miR-195 and so on, which showed that miRNAs isolated from LCEXL1210 were differentially expressed with those from the parental cells. Some differentially expressed miRNAs from LCEXL1210 were verified by real-time PCR, and then were analyzed by GO database, which demonstrated that these highly expressed miRNAs participated in the processes of various biological function and signal transduction. Conclusions MiRNAs isolated from LCEXL1210 show a high similarity to miRNAs isolated from L1210 cells, whereas of which one-third are specific. The highly expressed miRNAs participate in the processes of various biological function and signal transduction.

关 键 词：白血病 外泌体 基因本体 

分 类 号：R733.7[医药卫生—肿瘤]