MicroRNA PC-5p-1090 Inhibits Cell Proliferation By Targeting MARCKS

To explore the biological functions of low-copy microRNA PC-5p-1090(microRNA PC-5p-1090)that was newly discovered in antler velvet of red deer(Cervus elaphus)at rapid growth stage.In the research,MARCKS(myristoylated alanine-rich C-kinase substrate)which was membrane-binding protein involved in various signaling pathways and substrate of the protein kinase C(PKC)was used for functional validation as the predictive target of microRNA PC-5p-1090.The regulative effect of microRNA PC-5p-1090 on MARCKS was studied by luciferase reporter system and western blot.At the same time,cell proliferation assay was used to study the effect of microRNA PC-5p-1090 on cell physiological functions.The results showed that:(1)luciferase expression was significantly decreased when microRNA PC-5p-1090 mimics were co-transfected with wild-type MARCKS 3'UTR reporter construct in 293T cell line,while the luciferase expression levels of other control groups showed no significant changes.(2)when microRNA PC-5p-1090 mimics and NC mimics were separately transfected into Huh-7 cell line,western blot and cell proliferation assays indicated that the expression of endogenous MARCKS protein in microRNA PC-5p-1090 mimics group decreased significantly and Cell proliferation was inhibited.Studies had shown that:MicroRNA PC-5p-1090 was biologically functional;MicroRNA PC-5p-1090 specifically targeted to MARCKS 3'UTR and negatively regulated the expression of endogenous MARCKS gene,thus inhibiting cell proliferation.MicroRNA PC-5p-1090 regulatory mechanism to MARCKS was evolutionarily conservative.