LncRNA-599554 sponges miR-15a-5p to contribute inductive ability of dermal papilla cells through positive regulation of the expression of Wnt3a in cashmere goat

BACKGROUND: Long non-coding RNAs (lncRNAs), as post-transcriptional regulators, were thought to function in the inductive property of dermal papilla cells (DPCs) in cashmere goat. Previously, lncRNA-599554 was identified in secondary hair follicle (SHF) of cashmere goat, but its functional significance is unknown. RESULTS: In the present investigation, we verified that lncRNA-599554 had significantly higher expression at the anagen dermal papilla of cashmere goat SHF than that at telogen. Based on overexpression and knockdown techniques, we found that lncRNA-599554 contributes the inductive property of DPCs of cashmere goat, which was assessed by detecting the changes in the expression of several typical indictor genes in DPCs including ET-1, SCF, Versican, ALP, Lef1 and Ptc-1. Based on RNA pull-down assay, we verified that lncRNA-599554 directly interacted with chi-miR-15a-5p. Also, we showed that lncRNA-599554 positively regulated the Wnt3a expression in DPCs but which did not appear to involve its modulating of promoter methylation. Based on the use of Dual-luciferase reporter assays, our data indicated that lncRNA-599554 regulated the Wnt3a expression through chi-miR-15a-5p-mediated post-transcriptional level. CONCLUSIONS: We showed that lncRNA-599554 contributes the inductive property of DPCs in cashmere goat which might be achieved through sponging chi-miR-15b-5p to promote the Wnt3a expression. The results from the present investigation provided a novel insight into the functional mechanism of lncRNA-599554 in the SHF regeneration of cashmere goat along with the formation and growth of cashmere fiber.

miR-15a-5p inhibits proliferation and migration of human hepatocellular carcinoma SMMC-7721 cells / 中国病理生理杂志

AIM:To observe the influence of high expression of miR-15a-5p on the proliferation and migration of human hepatocellular carcinoma SMMC-7721 cells.METHODS: The miR-15a-5p oligonucleotide , which was recon-structed with additional restriction sites of EcoRⅠand HindⅢ, was chemically synthesized and confirmed by sequencing . The miR-15a-5p eukaryotic expression system was constructed by pcDNA 6.2-GW/Em-GFP-pre-miR-15a-5p plasmid.The miR-15a-5p was transfected into the SMMC-7721 cells transiently by plasmid , and quantified by quantitative real-time PCR at the mRNA level.The cell viability was measured by CCK-8 assay, and the living cell counting was performed by the method of Trypan blue exclusion .The migration ability of the SMMC-7721 cells with high expression of miR-15a-5p was de-tected by wound healing test .RESULTS: The sequence of miR-15a-5p oligonucleotide 100% matched the designed se-quence.Compared with control group , the miR-15a-5p expression was increased significantly (P<0.05).The viability, the living cell number and the migration ability of the SMMC-7721 cells were decreased in high expression of miR-15a-5p group with statistically significant difference (P<0.05).CONCLUSION: The abilities of proliferation and migration in human hepatocellular carcinoma SMMC-7721 cells are decreased by high expression of miR-15a-5p.