黄曲霉毒素B1诱导的恶性转化肝细胞miRNA表达谱的变化

Differentially expressed miRNA profile of transformed hepatic cells induced by aflatoxin B1

目的:检测黄曲霉毒素B1(aflatoxin B1,AFB1)诱导的恶性转化肝L02细胞(L02T)的miRNA表达谱,寻找差异表达的miRNA。方法:以含有AFB1的培养液多次间歇性染毒L02细胞获得转化细胞L02T,通过miRNA芯片技术检测和分析对照L02和转化L02T细胞的miRNA表达谱;用实时荧光定量PCR方法对芯片结果加以验证;采用TargetScan软件预测miRNA可能调控的靶基因。结果:获得2组细胞856个miRNA的表达谱,在25个表达差异显著的miRNA中,15个表达上调,10个表达下调;用定量RT-PCR对芯片结果中表达差异的miR-320a、miR-638和miR-98进行验证,并对其中上调显著的miR-638进行生物信息学分析,预测到4个与肝癌相关的潜在靶基因。结论:在AFB1诱导的恶性转化肝L02细胞中筛查出25个表达差异显著的miRNA,差异表达的miRNA可能在细胞恶性转化过程中起重要作用。

OBJECTIVE：To identify the differentially expressed miRNAs in malignantly transformed L02 cells（L02T） induced by aflatoxin B1（AFB1）,we performed miRNA microarray on both control cells（L02） and L02T cells.METHODS：L02T cells were obtained after exposure to aflatoxin B1（AFB1） several times.The comparison of differential expression profiles between L02 and L02T cells was performed and analyzed.The differentially expressed miRNAs were then selected for validation by semi-quantitative real-time PCR.The targets of miRNAs were predicted by TargetScan program.RESULTS：856 human miRNAs were analyzed by miRNA microarray.25 differentially expressed miRNAs were found in L02T cells compared with L02 cells,with 15 upregulated and 10 downregulated.Among these miRNAs,miR-320a,miR-638 and miR-98 were vahdated by quantitative real-time PCR.Moreover,predicted by TargetScan program,we identified 4 genes that were potentially targeted by miR-638. These genes might be associated with hepatic cell transformation induced by AFB1.CONCLUSION：25 differentially expressed miRNAs were identified in AFB1—transformed L02T cells.These miRNAs may play important roles in the process of malignant transformation of human cells.