title: Global identification of miR-373-regulated genes in breast cancer by quantitative proteomics.
abstract: Although microRNAs (miRNAs) have been reported to play an important role in carcinogenesis, their molecular mechanism remains largely unknown because of our limited understanding of miRNA target genes. miR-373 was found to be capable of promoting breast cancer invasion and metastasis, but only a target gene was experimentally identified on the basis of mRNA expression analysis. In this study, we used SILAC-based quantitative proteomics to globally identify the genes regulated by miR-373. Totally, 3666 proteins were identified, and 335 proteins were found to be regulated by miR-373. Among the 192 proteins that were downregulated by miR-373, 27 (14.1%) were predicted to have at least one potential match site at their 3'-UTR for miR-373 seed sequence. However, miR-373 did not affect the mRNA level of the five selected candidate targets, TXNIP, TRPS1, RABEP1, GRHL2 and HIP1, suggesting that the protein expressions were regulated by miR-373 via translational inhibition instead of mRNA degradation. Luciferase and mutation assays validated that TXNIP and RABEP1 were the direct target genes of miR-373. More than 30 proteins reported to be involved in cancer invasion and metastasis were found to be regulated by miR-373 in breast cancer for the first time.
authors.affiliation: Institute of Life and Health Engineering, and National Engineering and Research Center for Genetic Medicine, Jinan University, Guangzhou, P. R. China. tgryan@jnu.edu.cn
authors.forename: Guang-Rong
authors.initials: GR
authors.lastname: Yan
authors.forename: Song-Hui
authors.initials: SH
authors.lastname: Xu
authors.forename: Zi-Lu
authors.initials: ZL
authors.lastname: Tan
authors.forename: Langxia
authors.initials: L
authors.lastname: Liu
authors.forename: Qing-Yu
authors.initials: QY
authors.lastname: He
mesh_terms: D020413:3' Untranslated Regions; D001483:Base Sequence; D001665:Binding Sites; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D002465:Cell Movement; D030541:Databases, Genetic; D015536:Down-Regulation; D005260:Female; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D055106:Genome-Wide Association Study; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D009362:Neoplasm Metastasis; D040901:Proteomics / Q000379:methods*; D020871:RNA Stability; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D020413:3' Untranslated Regions; D002352:Carrier Proteins; D035683:MicroRNAs; C097227:RABEP1 protein, human; D012333:RNA, Messenger; C486715:TXNIP protein, human; D033921:Vesicular Transport Proteins; D008156:Luciferases