title: Screening for miRNAs and their potential targets in response to TGF-β1 based on miRNA microarray and comparative proteomics analyses in a mouse GC-1 spg germ cell line.
abstract: Transforming growth factor-β1 (TGF-β1) is a member of the TGF-β superfamily that performs a number of cellular functions and shows differential activity at different testicular developmental stages. In the present study, we investigated the effects of exogenous TGF-β1 on global microRNA (miRNA or miR) expression profiles by miRNA microarray analysis and the alterations in protein profiles by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/TOF MS) in a mouse GC-1 spg germ cell line. A total of 24 differentially expressed miRNAs, including 7 upregulated and 17 downregulated miRNAs were identified. The results obtained by the RT-qPCR analysis of 10 selected differentially expressed miRNAs were in accordance with those obtained by miRNA microarray analysis. In addition, 11 differentially expressed proteins, including 3 upregulated and 8 downregulated proteins were identified through MS-based comparative proteomics analysis. Bioinformatics analysis predicted that peptidyl‑prolyl isomerase A (PPIA) and nucleoside diphosphate kinase B (NDKB) are targets of miR-149 and miR-199a-3p, respectively in response to the stimulation of mouse GC-1 spg germ cells with TGF-β1. RT-qPCR revealed that the expression levels of these miRNAs showed an opposite trend in response to stimulation with TGF-β1. In conclusion, we identified some important miRNAs and proteins as possible targets involved in TGF-β1 signaling. Our data suggest the existence of a TGF-β1‑miR‑149-PPIA or TGF-β1-miR-199a-3p-NDKB pathway in GC-1 spg cells. Further studies are warranted to ascertain the role of these miRNAs in spermatogenesis.
authors.affiliation: Department of Life Science, College of Biology, Hunan University, Changsha, Hunan 410082, P.R. China.
authors.forename: Zhuoxian
authors.initials: Z
authors.lastname: Rong
authors.affiliation: Department of Life Science, College of Biology, Hunan University, Changsha, Hunan 410082, P.R. China.
authors.forename: Dan
authors.initials: D
authors.lastname: Li
authors.affiliation: Department of Life Science, College of Biology, Hunan University, Changsha, Hunan 410082, P.R. China.
authors.forename: Xiaowen
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Life Science, College of Biology, Hunan University, Changsha, Hunan 410082, P.R. China.
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Department of Life Science, College of Biology, Hunan University, Changsha, Hunan 410082, P.R. China.
authors.forename: Daobing
authors.initials: D
authors.lastname: Wu
authors.affiliation: Department of Life Science, College of Biology, Hunan University, Changsha, Hunan 410082, P.R. China.
authors.forename: Xuanming
authors.initials: X
authors.lastname: Liu
mesh_terms: D000818:Animals; D001665:Binding Sites; D019295:Computational Biology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D005854:Germ Cells / Q000187:drug effects* / Q000378:metabolism*; D051379:Mice; D035683:MicroRNAs / Q000737:chemistry / Q000235:genetics*; D020543:Proteome; D040901:Proteomics; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics*; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*
chemical_list: D035683:MicroRNAs; D020543:Proteome; D012333:RNA, Messenger; D053773:Transforming Growth Factor beta1