Expression of microRNA-125a in psoriasis vulgaris lesions and its effect on the proliferation of HaCaT cells
10.35541/cjd.20200480
- VernacularTitle:microRNA-125a在寻常型银屑病皮损中的表达及其对HaCaT细胞增殖的影响
- Author:
Fang SU
1
;
Liang JIN
;
Wei LIU
;
Hong CAI
;
Xiaodong SUN
;
Xiaojie SUN
;
Guijuan XU
;
Qiang WANG
Author Information
1. 沈阳市第七人民医院皮肤科 110003
- From:
Chinese Journal of Dermatology
2020;53(11):886-890
- CountryChina
- Language:Chinese
-
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.
