Chinese Journal of Dermatology ›› 2021, Vol. 54 ›› Issue (6): 499-503.doi: 10.35541/cjd.20200707

• Original Articles • Previous Articles     Next Articles

Mechanisms underlying microRNA-125a-mediated inhibition of proliferation of HaCaT cells by targeting the interleukin 23 receptor signaling pathway: a preliminary study

Su Fang1, Jin Liang2, Li Hao1, Ding Yingjie1, Sun Xiaojie1, Sun Xiaodong1, Liu Wei2, Xu Guijuan1, Wang Qiang1, Liu Yongbin1   

  1. 1Department of Dermatology, The Seventh People′s Hospital of Shenyang, Shenyang 110003, China; 2Department of Dermatology, Air Force Medical Center, Beijing 100142, China
  • Received:2020-07-13 Revised:2020-12-14 Online:2021-06-15 Published:2021-05-31
  • Contact: Liu Wei; Liu Yongbin;
  • Supported by:
    Science and Technology Planning Project of Liaoning Province(2019?ZD?0975); Young and Middle?aged Scientific and Technological Innovation Talent Support Program of Shenyang(RC200417)

Abstract: 【Abstract】 Objective To explore the mechanism underlying microRNA (miR)-125a-mediated inhibition of proliferation of keratinocytes. Methods After 24-hour pretreatment with interleukin (IL)-23, human HaCaT keratinocytes were divided into miR-125a group and miR-NC group transfected with a miR-125a overexpression plasmid and a control plasmid, respectively. Cell counting kit-8 (CCK8) assay was performed to evaluate the proliferative ability of HaCaT cells in the two groups at 0, 24, 48 and 72 hours after transfection, real-time fluorescence-based quantitative PCR to determine the mRNA expression of miR-125a and IL-23 receptors (IL-23R) in the two groups 24 hours after transfection, and Western blot analysis to determine the protein expression of IL-23R, Janus kinase 2 (JAK2), protein kinase B (AKT) and phosphorylated AKT (p-AKT) in the two groups 48 hours after transfection. Dual-luciferase reporter assay was performed to verify the targeting relationship between miR-125a and IL-23R. Comparison of means between two groups was carried out by using t test, and changes in the proliferative ability of HaCaT cells over time were evaluated by using repeated measures analysis of variance. Results After plasmid transfection, the relative expression of miR-125a was significantly higher in the miR-125a group (6.377 ± 0.745) than in the miR-NC group (0.700 ± 0.222; t = 7.305, P = 0.002). At 0, 24 and 48 hours after transfection, there was no significant difference in cellular proliferative ability between the miR-125a group and the miR-NC group (t = 0.663, 0.623 and 1.930, respectively, all P > 0.05); at 72 hours after transfection, the cellular proliferative ability was significantly lower in the miR-125a group than in the miR-NC group (t = 4.407, P < 0.05). The IL-23R mRNA expression was significantly lower in the miR-125a group than in the miR-NC group (t = 3.082, P < 0.05). Compared with the miR-NC group, the miR-125a group showed significantly decreased protein expression of IL-23R, JAK2 and p-AKT (t = 11.715, 6.996, 12.424, P < 0.001, = 0.002, < 0.001, respectively). Dual-luciferase reporter assay showed targeted binding of miR-125a to IL-23R. Conclusion MiR-125a may inhibit the proliferation of keratinocytes by negatively regulating the IL-23R/JAK2/AKT signaling pathway.

Key words: Psoriasis, Keratinocytes, MicroRNAs, Cell proliferation, Interleukin-23, MicroRNA-125a