Chinese Journal of Dermatology ›› 2020, Vol. 53 ›› Issue (11): 886-890.doi: 10.35541/cjd.20200480

• Original Articles • Previous Articles     Next Articles

Expression of microRNA-125a in psoriasis vulgaris lesions and its effect on the proliferation of HaCaT cells

Su Fang1, Jin Liang2, Liu Wei2, Cai Hong2, Sun Xiaodong1, Sun Xiaojie1, Xu Guijuan1, Wang Qiang1   

  1. 1Department of Dermatology, The Seventh People′s Hospital of Shenyang, Shenyang 110001, China; 2Department of Dermatology, Air Force Medical Center, Beijing 100142, China
  • Received:2020-05-18 Revised:2020-09-17 Online:2020-11-15 Published:2020-11-03
  • Contact: Liu Wei; Wang Qiang E-mail:lwei5811@126.com; swepco@126.com
  • Supported by:
    Science and Technology Planning Project of Liaoning Province (2019-ZD-0975)

Abstract: 【Abstract】 Objective To investigate the correlation between microRNA-125a(miR-125a) expression and inflammatory cytokine levels in skin lesions of patients with psoriasis vulgaris, and to evaluate the effect of miR-125a on the proliferation of a human immortalized keratinocyte cell line HaCaT. Methods Totally, lesional and adjacent non-lesional skin tissues were collected from 40 patients with psoriasis vulgaris in the Seventh People′s Hospital of Shenyang from 2017 to 2018, and real-time fluorescence-based quantitative reverse transcription PCR was performed to determine the expression of miR-125a in the skin tissues, as well as the mRNA expression of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, and IL-17 in the lesional skin tissues. HaCaT cells were divided into 4 groups to be transfected with a miR-125a overexpression plasmid (miR-125a overexpression group), an overexpression control plasmid (overexpression control group), a miR-125a interference plasmid (miR-125a interference group) and an interference control plasmid (interference control group), respectively. Cell counting kit-8 (CCK8) assay was performed to assess the proliferative ability of HaCaT cells in the groups at 0, 24, 48, 72 hours after transfection, and double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) to detect the levels of TNF-α, IL-1β, IL-6 and IL-17 in the culture supernatant of HaCaT cells. Spearman rank correlation test was used for correlation analysis, and t test for the comparison of means between two groups. Results The relative expression of miR-125a was significantly lower in the lesional skin tissues (expressed as 2-ΔΔCt, 0.389 ± 0.354) than in the non-lesional skin tissues (1.106 ± 0.396, t = 7.717, P < 0.001) in patients with psoriasis vulgaris. The expression of miR-125a was negatively correlated with the mRNA expression of TNF-α, IL-1β and IL-17 in psoriatic lesions (r = -0.447, -0.424, -0.436, all P < 0.01). Immediately and 24 hours after transfection with the plasmids, there was no significant difference in the cell proliferative ability between the miR-125a overexpression group and overexpression control group (t = 0.282, 1.445, respectively, both P > 0.05), or between the miR-125a interference group and interference control group (t = 0.120, 1.543, respectively, both P > 0.05). Forty-eight and 72 hours after the transfection, the cell proliferative ability was significantly lower in the miR-125a overexpression group than in the overexpression control group (t = 3.222, 4.563, respectively, both P < 0.05), but significantly higher in the miR-125a interference group than in the interference control group (t = 3.036, 3.269, respectively, both P < 0.05). In addition, the miR-125a overexpression group showed significantly decreased levels of TNF-α and IL-1β compared with the overexpression control group (t = 4.318, 3.813, respectively, both P < 0.05). Conclusions MiR-125a is lowly expressed in skin lesions of patients with psoriasis vulgaris. MiR-125a can inhibit the proliferation of keratinocytes, and may play a protective role in the occurrence and development of psoriasis.

Key words: Psoriasis, MicroRNAs, Keratinocytes, Cell proliferation, Tumor necrosis factor-alpha, Interleukin-1beta, Interleukin-6, Interleukin-17