肝癌干细胞差异表达miRNA筛选及鉴定.docVIP

肝癌干细胞差异表达miRNA筛选及鉴定作者单位：224000 江苏省盐城市第一人民医院 通讯作者：周广军 【摘要】 目的 培养卵圆细胞、肝癌干细胞样细胞，筛选肝癌干细胞差异表达的microRNA并进行验证。方法 体外分离培养卵圆细胞，肝癌干细胞样细胞，通过microRNA芯片检测差异表达的microRNA，对所得到的差异表达显著的microRNA进行Northern验证。结果 表达差异显著的microRNA共有17个，经Northern杂交验证，其中6个差异显著的microRNA，发现hsa-let-7f-2、hsa-miR-199a-3p、hsa-miR-122在肝癌干细胞中的表达较卵圆细胞明显升高。结论 hsa-let-7f-2、hsa-miR-199a-3p、hsa-miR-122在肝癌干细胞中的表达较卵圆细胞明显升高，可以进一步验证其在肝癌干细胞中的功能。 【关键词】 miRNAs； 肝癌干细胞； 芯片谱 Screen and identification of differentially expressed miRNAs in liver cancer stem cells and oval cells ZHOU Guang-jun，SHI Kun，ZHA Wen-zhang，MU Xiang-ming. The Fist People’s Hospital of Yancheng City, Yancheng 224000,China 【Abstract】 Objective To determine differentially expressed miRNAs of liver cancer stem cells and oval cells.Methods Liver cancer stem cells and oval cells were isolated from the livers of human cancer patients. The difference in miRNAs expression between liver cancer stem cells and oval cells was detected by both microarrays and Northern blot.Results Forteen miRNAs in liver cancer stem cells were expressed 1.5 fold above the levels in oval cells, three miRNAs in liver cancer stem cells were expressed 1.5 fold below the levels in oval cells. The expression of six selected miRNAs was further validated by Northern blot and three were confirmed.Conclusion The expression of a number of miRNAs in liver cancer stem cells and oval cells may suggest the potential roles of these miRNAs and applicability as diagnostic and prognostic signatures in liver cancer. 【Key words】 MicroRNAs; Liver cancer stem cells; Microarrays 肿瘤干细胞（tumor stem cells,TSC）是近年来实体肿瘤研究的热点，对实体瘤认识产生了重大影响。肝癌、脑恶性肿瘤、乳腺癌、前列腺癌等恶性肿瘤中都发现了TSC，并且其表面标记特征与生长特征等也得到一定阐述［1］。TSC具备高度增殖能力与自我更新能力，也具备多向分化的潜能，是一种特殊类型的干细胞。TSC通过不均一分裂，一个TSC分裂形成一个新的TSC，和另一个可最终分化为包括肿瘤细胞在内的各种细胞的子细胞，其结果是维持TSC数目稳定并产生肿瘤。TSC的特性包括极强的致瘤能力与自我更新并多向分化，并且有高耐药的特征。目前认为，TSC来源于正常成体干细胞［1］。 肝癌是TSC研究的热点之一。肝癌也越来越被设想为一种干细胞疾病。近年来，本课题组人员从大鼠肝癌中分离、培养并纯化出一些有干细胞特性的细胞，有强的致瘤能力，与TSC比较接近，并且对这些细胞的增殖能力，转移能力，细胞生长特征，细胞周期分析等生物学特征也进行了初步的研