Proteomic identification of microRNA-122a target proteins in hepatocellular carcinoma.
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.
Diao
Shu
S
Department of Chemistry, The Chinese University of Hong Kong, Hong Kong.
Zhang
Jin-fang
JF
Wang
Hua
H
He
Ming-liang
ML
Lin
Marie Chia-mi
MC
Chen
Yangchao
Y
Kung
Hsiang-fu
HF
D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D054464:Peroxiredoxins / Q000235:genetics / Q000378:metabolism*; D020543:Proteome / Q000032:analysis*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
C531907:MIRN122 microRNA, human; D035683:MicroRNAs; D009363:Neoplasm Proteins; D020543:Proteome; C086065:PRDX2 protein, human; D054464:Peroxiredoxins