Promoting Effect of MicroRNA-9-1 on Differentiation of Epidermal Stem Cells into Neural Cells in Rats / 现代生物医学进展

ABSTRACT

Objective:To investigate the role of MicroRNA-9-1 in inducing epidermal stem cells(ESCs) differentiation into neurons.Methods:The lentiviral of MicroRNA-9-1 was constructed and transfected into rats epidermal stem cells.The experiment was divided into transfected group,non-transfected group and the negative control group.The β-mercaptoethanol was as an inducer for triggering the ESCs to differentiate into neurons.The GFP fluorescence expression of epidermal stem cells after transfection was observed under inverted fluorescence microscope.The protein and mRNA expression level of microtuble-associated protein 2 (MAP-2) was detected by immunocytochemical method and RT-PCR,respectively.Results:The result of Positive clone PCR confirmed successful construction of MicroRNA-9-1 in rats.Transfection after 48 h,the expressing of GFP fluorescence at peak in transfected group,and transfection efficiency reached (85.6+1.9)％.Most ESCs differentiated into neurons in transfected group after β-mercaptoethanol induction 7 h,and the effect was significantly better than non-transfected group and the negative control group.The protein ((87.3± 0.6)％) and mRNA (about twice over) expression levels of MAP-2 in transfected group was higher than those in non-transfected group and the negative control group (P＜0.05).Conclusion:The lentiviral of MicroRNA-9-1 has high transfection efficiency in rats ESCs,and could promoted ESCs differentiate into neurons under β-mercaptoethanol induced.