Abstract
Human amnion mesenchymal stem cells (hAMSCs) are promising sources of stem cells in regenerative medicine. The migration stimulated by cytokines is critical for mesenchymal stem cells (MSCs)-based cytotherapy, while the regulatory mechanisms of EGF (epidermal growth factor)-induced hAMSC migration are largely unclear. Here, a novel miRNA N-72 (GenBank accession number: MH269369) has been discovered, and its function on EGF-induced migration in hAMSCs was investigated. High-purity hAMSCs were isolated and cultured in vitro, which were characterized by flow cytometry and trilineage differentiation. The N-72 located on chromosome three was conserved, and pri-N-72 owned the ability to form a stem-loop secondary structure, which was predicated by bioinformatic programs. The expression of mature N-72 was verified in several human cells including hAMSC by real-time PCR. In EGF-stimulated hAMSC, N-72 showed a significant reduction in a PI3K and p38 MAPK-dependent manner, and N-72 mimics transfection-inhibited EGF-induced migration, which was verified by scratch assay and transwell assay. Further, the predicated target gene MMP2 was proved to be a direct target of N-72 via luciferase reporter assay, real-time PCR, and Western blotting. The results that MMP2 silencing repressed hAMSC migration suggested MMP2 as a functional downstream target of N-72. In summary, we have discovered the novel N-72, and it was crucial for EGF-induced migration by targeting MMP2 in hAMSCs.
Highlights
Cytotherapy using mesenchymal stem cells (MSCs) provides novel strategies for the treatment of various diseases, and MSCs have been isolated and expanded in vitro from different tissues such as bone marrow, adipose tissue, and amnion tissue [1,2,3]
epidermal growth factor factor (EGF) could enhance therapeutic potentials including the motility of MSCs, and recent studies demonstrated that human amnion mesenchymal stem cells show great differentiation and proliferation potential, as well as other remarkable features that could serve as an outstanding alternative source of stem cells in regenerative medicine [8], while the regulatory mechanisms of EGF-induced hAMSC migration are unclear
In hAMSC, NCBI BLAST and ClustalW among different species, the results showed that N-72 is conserved in the expression
Summary
Cytotherapy using mesenchymal stem cells (MSCs) provides novel strategies for the treatment of various diseases, and MSCs have been isolated and expanded in vitro from different tissues such as bone marrow, adipose tissue, and amnion tissue [1,2,3]. The migration of MSCs to diseased regions is critical for their clinical application. This migration could be stimulated by the cytokines and chemokines secreted from diseased regions such as TGF-β, VEGF, and EGF (epidermal growth factor) [4,7].
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