Department of Chemistry, The Chinese University of Hong Kong, Hong Kong.
Proteomics. 2010 Oct;10(20):3723-31. doi: 10.1002/pmic.201000050.
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.
微小 RNA-122a(miR-122a)是一种肝脏特异性 miRNA,在肝细胞癌(HCC)中经常下调。miR-122a 及其在 HCC 中的靶标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前 miR-122a(Lenti-miR-122a)的慢病毒载体。Lenti-miR-122a 抑制 HCC 细胞体外生长并诱导细胞凋亡。我们采用蛋白质组学分析来鉴定 miR-122a 的靶蛋白。总共鉴定出 10 种在感染 Lenti-miR-122a 的 HCC 细胞中差异表达的蛋白质。其中,PRDXII 的下调通过 Western blot 得到验证。通过生物信息学分析,在 PRDXII mRNA 的 5'-UTR 中鉴定出 miR-122a 的可预测靶位点。荧光素酶报告基因检测证实了 miR-122a 对 PRDXII 5'-UTR 的调控。总之,PRDXII 被鉴定为 miR-122a 的新靶标。