Identification of the receptor tyrosine kinases (RTKs)-oriented functional targets of miR-206 by an antibody-based protein array

Yazhou Cui; Shuyang Xie; Jing Luan; Xiaoyan Zhou; Jinxiang Han

doi:10.1016/j.febslet.2015.06.026

Identification of the receptor tyrosine kinases (RTKs)-oriented functional targets of miR-206 by an antibody-based protein array

FEBS Lett. 2015 Jul 22;589(16):2131-5. doi: 10.1016/j.febslet.2015.06.026. Epub 2015 Jul 2.

Authors

Yazhou Cui¹, Shuyang Xie², Jing Luan¹, Xiaoyan Zhou¹, Jinxiang Han³

Affiliations

¹ Shandong Academy of Medical Sciences, Shandong Medical Biotechnological Center, Key Laboratory for Biotech Drugs of the Ministry of Health, China.
² Department of Biochemistry and Molecular Biology, Binzhou Medical University, YanTai, Shandong, China.
³ 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.

PMID: 26143374
DOI: 10.1016/j.febslet.2015.06.026

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.

Keywords: Antibody array; MicroRNA; Proteomic; Target gene; miR-206.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antibodies, Phospho-Specific / metabolism*
Antibody Specificity
Binding Sites
Cell Line, Tumor
Computational Biology
Eukaryotic Initiation Factor-4E / antagonists & inhibitors*
Eukaryotic Initiation Factor-4E / genetics
Eukaryotic Initiation Factor-4E / metabolism
Genes, Reporter
Humans
MicroRNAs / metabolism*
Mutant Proteins / antagonists & inhibitors
Mutant Proteins / metabolism
PAX3 Transcription Factor
Paired Box Transcription Factors / antagonists & inhibitors
Paired Box Transcription Factors / genetics
Paired Box Transcription Factors / metabolism
Phosphorylation
Promoter Regions, Genetic*
Protein Array Analysis
Protein Processing, Post-Translational
Proteomics / methods
Proto-Oncogene Proteins c-met / antagonists & inhibitors*
Proto-Oncogene Proteins c-met / genetics
Proto-Oncogene Proteins c-met / metabolism
RNA Interference*
Recombinant Proteins / chemistry
Recombinant Proteins / metabolism
Sorting Nexins / antagonists & inhibitors
Sorting Nexins / genetics
Sorting Nexins / metabolism

Substances

Antibodies, Phospho-Specific
Eukaryotic Initiation Factor-4E
MIRN206 microRNA, human
MicroRNAs
Mutant Proteins
PAX3 Transcription Factor
PAX3 protein, human
Paired Box Transcription Factors
Recombinant Proteins
SNX2 protein, human
Sorting Nexins
MET protein, human
Proto-Oncogene Proteins c-met