大鼠肝卵圆细胞向肝癌细胞分化过程中microRNA的变化  

MicroRNA differential expression profile during differentiation of rat oval cells toward hepatocellular carcinoma

作　　者：许荣华[1] 何冬雷[1] 孟津[1] 夏立平[1] 王健[2] 

机构地区：[1]海南医学院附属医院胃肠肿瘤外科,海南海口570102 [2]南方医科大学附属花都医院普外科,广东广州510800

出　　处：《中国现代医学杂志》2014年第7期1-6,共6页China Journal of Modern Medicine

基　　金：海南省自然科学基金项目(No:309063);2011年广东省第三批科技计划项目(No:201197)

摘　　要：目的利用生物芯片技术分析大鼠肝卵圆细胞分化为肝癌细胞过程中微小RNA(microRNA,miRNA)的表达变化,筛选调控分化的microRNAs,探讨肝癌发生的起源机制。方法用化学致癌剂3’-Me-DAB诱发SD大鼠肝卵圆细胞增生,采用密度梯度离心法分离、纯化肝卵圆细胞,行体外培养使其恶性分化并进行鉴定。提取分化过程中的microRNA进行microRNA基因芯片杂交,获得卵原细胞恶性分化过程中microRNA表达谱。同时利用种植模型及大鼠Y染色体特异性PCR技术,验证卵圆细胞可恶性分化为肝癌细胞。结果①激光扫描共聚焦显微镜显示,肝卵圆细胞胞浆和胞膜表达干细胞标志Thy-1及C-kit;②通过基因微阵列分析,卵原细胞恶性分化过程中有22个miRNA的改变显著,其中19个表达上调,3个表达下调;③卵原细胞种植模型的大鼠肝癌组织具有SRY基因的电泳条带。结论特定microRNA可能对大鼠肝卵圆细胞分化为肝癌细胞过程起到关键作用。[ Objective ] To explore the expression profile of microRNAs during differentiation of rat hepatic oval cells（HOCs） toward hepatocellular carcinoma, so as to provide a basis for further exploring the role of miRNAs in the molectdar mechanism that leads to the occurrence and development of live cancer. [Methods] Proliferation of rat HOCs was induced by chemical carcinogen, 3＇-Me-DAB. HOCs were isolated By Percoll density gradient eentrifugation method, followed by continuous cultivation in vitro. Total RNA was extracted from HOCs during cells differentiation, and mieroRNA was isolated from the total RNA. Mieroarray analysis of mieroRNA expression was performed to detect the different expression levels of microRNA among the indicated time points. Rats＇ Y chromosome specific polymerase chain reaction technique was used to find out the evidence that HOCs could differentiated into liver cancer cells. [ Results ] Laser scanning confoca] microscopy indicated positive expression of stem cells markers Thy-1 and C-kit in cytoplasm and membrane of HOCs. A total of 1 210 miRNAs were identified and among them 22 were differentially expressed（P 〈0.05, fold ehange≥2）, including 19 up-regulated and 3 down-regulated ones. In vitro HOCs implantation model showed the same length of positive bands after electrophoresis designed by the spe cific primer of rat SRY gene. [Conclusion] Some specific mieroRNAs play a key role in the course that HOCs differentiate into liver cancer cells.

关 键 词：肝卵圆细胞 分化 MICRORNA 微小RNA芯片 

分 类 号：R-332[医药卫生]