MiR-506-3p regulates autophagy and proliferation in post-burn skin fibroblasts through post-transcriptionally suppressing Beclin-1 expression

Min Shi, Xiaoming Zong, Lei Chen, Xiaobo Guo, Xinqiang Ding
In Vitro Cellular & Developmental Biology. Animal 2020 August 4
MicroRNAs (miRNAs) is involved in diverse biological processes of cells including dermal fibroblasts that contributed to wound healing and resulted in keloid scarring. MiR-506-3p has been identified as a tumor suppressor or oncogene in fibroblasts of various cancers, while the role of miR-506-3p in regulating functions of post-burn dermal fibroblasts is poorly known. In this study, miR-506-3p was confirmed to be significantly downregulated in burned tissues and heat-stimulated dermal fibroblasts. Expression levels of autophagy-related proteins suggested thermal stimulus promoting the autophagy in dermal fibroblasts. Then, miR-506-3p inhibition enhanced cell proliferation and cell cycle process in dermal fibroblasts after thermal stimulus, whereas overexpression of miR-506-3p showed the opposite effect. Western blot assay showed that inhibition of miR-506-3p resulted in the upregulation of the expression levels of LC3-II, ATG5, and structural protein collagen I, as well as the downregulation of p62. Marker proteins of intermolecular cross-links in collagen synthesis, including hydroxylysylpyridinoline (HP), lysinepyridine (LP), and lysyl hydroxylase 2 (LH2), were increased by miR-506-3p overexpression and decreased by miR-506-3p inhibition. Moreover, transfection with miR-506-3p mimic suppressed the proliferation and autophagy in heat-stimulated dermal fibroblasts in a dose-dependent manner. Subsequently, dual luciferase reporter gene assay demonstrated that Beclin-1 was a direct target of miR-506-3p, and reintroduction of Beclin-1 could antagonize the suppressive effect of miR-506-3p overexpression on fibroblast proliferation, autophagy, and the intermolecular cross-links in collagen synthesis. Taken together, our findings showed that miR-506-3p regulated autophagy and proliferation in post-burn skin fibroblasts through post-transcriptionally suppressing Beclin-1 expression.

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