Proteomic identification of microRNA-122a target proteins in hepatocellular carcinoma
Proteomic identification of microRNA-122a target proteins in hepatocellular carcinoma
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DOI：
10.1002/pmic.201000050
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发表时间：
2010-10-01
期刊：
影响因子：
3.4
通讯作者：
Kung, Hsiang-fu
中科院分区：
文献类型：
--
作者：
Diao, Shu;Zhang, Jin-fang;Kung, Hsiang-fu
microRNA-122 a（miR-122 a）是一种肝脏特异性miRNA，在肝细胞癌（HCC）中经常下调。miR-122 a及其靶点在HCC中的确切功能作用在很大程度上仍然未知。我们开发了一种用于表达pre-miR-122 a（Lenti-miR-122 a）的慢病毒载体。Lenti-miR-122 a在体外抑制HCC细胞生长并诱导凋亡。我们采用蛋白质组学分析来鉴定miR-122 a的靶蛋白。总共鉴定了在用Lenti-miR-122 a感染的HCC细胞中具有差异表达的十种蛋白质。其中，通过Western印迹验证了miR-122 a对过氧化物酶氧还蛋白2（PRDXII）的下调。使用生物信息学分析，在PRDXII mRNA的5 '-UTR中鉴定了miR-122 a的可预测靶位点。荧光素酶报告基因检测证实了miR-122 a对PRDXII 5 '-UTR的调控作用。总之，PRDXII被鉴定为miR-122 a的新靶点。
microRNA-122a (miR-122a) is a liver-specific miRNA that is frequently downregulated in hepatocellular carcinoma (HCC). The exact functional role of miR-122a and its target in HCC remain largely unknown. We developed a lentiviral vector for the expression of pre-miR-122a (Lenti-miR-122a). Lenti-miR-122a inhibited HCC cell growth and induced apoptosis in vitro. We employed proteomic profiling to identify the target proteins of miR-122a. In total, ten proteins with differential expression in HCC cells infected with Lenti-miR-122a were identified. Amongst them, downregulation of peroxiredoxin 2 (PRDXII) by miR-122a was validated by Western blotting. Using bioinformatics analysis, predictable target sites of miR-122a were identified in the 5'-UTR of PRDXII mRNA. Luciferase reporter assay confirmed the regulation of miR-122a on 5'-UTR of PRDXII. In conclusion, PRDXII was identified to be the new target of miR-122a.