title: Identification of the receptor tyrosine kinases (RTKs)-oriented functional targets of miR-206 by an antibody-based protein array.
abstract: This study demonstrated the feasibility and benefit of an antibody-based experimental approach to identify microRNA functional targets from hundreds of predicted genes using miR-206 as an example. Using a receptor tyrosine kinase (RTK) antibody array, we identified 7 phosphorylated RTKs that were significantly differentially regulated after miR-206-mimic transfection. We then focused on MET, the most varied RTK, and bioinformatically constructed a MET-centred network using computationally predicted miR-206 targets. Within this network, we analyzed two validated targets, PAX3 and SNX2, and one candidate target, EIF4E, may account for the inhibitory effect of miR-206 on MET phosphorylation. Luciferase and Western-blot assays indicated that EIF4E was a direct target of miR-206. This concept may also be applicable for other microRNAs and other antibody array platforms.
authors.affiliation: Shandong Academy of Medical Sciences, Shandong Medical Biotechnological Center, Key Laboratory for Biotech Drugs of the Ministry of Health, China.
authors.forename: Yazhou
authors.initials: Y
authors.lastname: Cui
authors.affiliation: Department of Biochemistry and Molecular Biology, Binzhou Medical University, YanTai, Shandong, China.
authors.forename: Shuyang
authors.initials: S
authors.lastname: Xie
authors.affiliation: Shandong Academy of Medical Sciences, Shandong Medical Biotechnological Center, Key Laboratory for Biotech Drugs of the Ministry of Health, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Luan
authors.affiliation: Shandong Academy of Medical Sciences, Shandong Medical Biotechnological Center, Key Laboratory for Biotech Drugs of the Ministry of Health, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Shandong Academy of Medical Sciences, Shandong Medical Biotechnological Center, Key Laboratory for Biotech Drugs of the Ministry of Health, China. Electronic address: jxhan9888@aliyun.com.
authors.forename: Jinxiang
authors.initials: J
authors.lastname: Han
mesh_terms: D045163:Antibodies, Phospho-Specific / Q000378:metabolism*; D000918:Antibody Specificity; D001665:Binding Sites; D045744:Cell Line, Tumor; D019295:Computational Biology; D039561:Eukaryotic Initiation Factor-4E / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D017930:Genes, Reporter; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D050505:Mutant Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D040081:Protein Array Analysis; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods; D019859:Proto-Oncogene Proteins c-met / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D058305:Sorting Nexins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
chemical_list: D045163:Antibodies, Phospho-Specific; D039561:Eukaryotic Initiation Factor-4E; C546087:MIRN206 microRNA, human; D035683:MicroRNAs; D050505:Mutant Proteins; D000072041:PAX3 Transcription Factor; C489409:PAX3 protein, human; D051761:Paired Box Transcription Factors; D011994:Recombinant Proteins; C513979:SNX2 protein, human; D058305:Sorting Nexins; C491743:MET protein, human; D019859:Proto-Oncogene Proteins c-met
keywords: Antibody array; MicroRNA; Proteomic; Target gene; miR-206