Effects of miR-124a overexpression on expression levels of TNF-α and IL-6 and cell cycle of J774. 1 cells / 吉林大学学报(医学版)

ABSTRACT

Objective; To explore the effects of miR-124a on the expression levels of tumor necrosis factor alpha (TNF-a) and interleukin-6 (IL-6) and cell cycle in the macrophage J774. 1 cells of the rheumatoid arthritis (RA) model mice, and to elucidate the mechanism of miR-124a in the pathogenesis of RA. Methods: The J774. 1 cells in logarithmic growth phage were obtained and uniformly inoculated in the petri dish with 1 × 106 mL-1, and there were 3 multiple holes in each group; transfection was carried out when the fusion degree of the cells reached 60%. The cells were divided into blank control group (without any treatment), empty vector group (transfected with adenovirus negative fluid) and miR-124a overexpression group (transfected with miR-124a adenovirus vector). The expression levels of TNF-α and IL-6 in supernatant of the cells in three groups were detected by ELISA 48 h after transfection. RT-PCR was used to detect the relative expression levels of TNF-α and IL-6 mRNA in J774. 1 cells 48 h after transfection and the changes of cell cycle were detected using flow cytometry. Results: The expression levels of TNF-α and IL-6 in supernatant of the cells in miR-124a overexpression group were lower than those in blank control group and empty vector group 48 h after transfection (P=0. 038, P=0. 042; P=0. 043, P=0. 044). The expression levels of TNF-α mRNA and IL-6 mRNA in miR-124a group were significantly lower than those in blank control group and empty vector group (P = 0. 001, P=0.002; P=0. 001, P=0. 003). The pecentage of cells in Gi phase in miR-124a overexpression group was higher than those in blank control group and empty vector group (P<0. 01); the percentages of cells in S phase and G2 phase were lower than those in blank control group and empty vector group (P<0.01). Conclusion: MiR-124a can decrease the expression levels of inflammatory cytokines after transfecting the macrophages J774. 1 cells, and inhibit the proliferation of cells. MiR-124a is a potential therapeutic target for the treatment of RA.