microRNA在两种不同来源的肝卵圆细胞中的差异表达

Differential expression profile of microRNAs in the hepatic oval cells derived from different origins

目的分离纯化两种不同模型来源的卵圆细胞(HOCs)并初步筛选差异表达的microRNA。方法采用2-乙酰氨基芴(2-AAF)和2/3肝切除(PH)建立SD大鼠HOCs增殖模型,用化学致癌剂3'-甲基-4-二甲基偶氮苯(3'-Me-DAB)诱发SD大鼠肝卵圆细胞增生模型;分离纯化并鉴定两种不同模型来源的大鼠肝卵圆细胞;用microRNA芯片技术检测HOCs/AAF与HOCs/DAB中microRNA的表达谱,筛选差异表达的microRNA;用实时荧光定量PCR方法对筛选出的部分差异microRNA进行验证。结果 1激光扫描共聚焦显微镜显示,两种肝卵圆细胞胞质和胞膜均表达干细胞标志Thy-1及C-kit。2microRNA芯片的结果显示,HOCs/DAB与HOCs/AAF比较,有10个microRNA表达上调,8个microRNA表达下调。3实时荧光定量PCR结果进一步证实mir-210、mir92b、mir-1281在HOCs/DAB中显著上调,而mir-181、mir-1228显著下调。结论两种不同模型来源的肝卵圆细胞存在着显著差异表达的microRNA,为研究肝癌细胞可能来源于肝卵圆细胞HOCs/DAB提供线索。

Objective To explore the microRNAs(miRNAs) differential expression profile in the hepatic oval cells(HOCs) which isolated and purified from two types of rat modes. Methods Wistar adult rats were fed by 2-acetaminofluorene(AAF) and were stimulated by partial hepatectomy to activate the proliferation of HOCs /AAF. HOCs /DAB response were stimulated in rats by treated with 3'-Me-DAB diet.The miRNAs expression difference between HOCs /AAF and HOCs /DAB was tested by microarray and selected several miRNAs candidates to further confirm by Q-PCR. Results 1Laser scanning confocal microscopy indicated positive expression of stem cells markers Thy-1 and Ckit in cytoplasm and membrane of HOCs /DAB,also in HOCs /AAF. 2Microarray suggested,by contrast with HOCs /AAF,10 miRNAs were up-regulated and 8 miRNAs were down-regulated in HOCs /DAB. 3Q-PCR further confirmed mir-210,mir92 b,mir-1281 up-regulated dramatically,but mir-181,mir-1228 down-regulated markedly between HOCs /DAB and HOCs /AAF. Conclusions The miRNA expression profiles of HOCs /DAB and HOCs /AAF are obtained. Liver cancer cells might be come from HOCs /DAB.