Journal of Jilin University(Medicine Edition) ›› 2022, Vol. 48 ›› Issue (3): 773-782.doi: 10.13481/j.1671-587X.20220327

• Research in clinical medicine • Previous Articles    

Inhibitory effect of silencing expression of DNMT3a on progression of psoriasis like cell cycle and cell proliferation and its mechanism

Yanbin PAN1,Jiaguang SU2,Meile TAN1,Meng YANG1,Wenfei TAN1,Yuxiu HUANG1,Shihao MENG1,Yaohui HUANG1,Jianqiang LIANG1,Xuefang SU1,Zichan HUANG1,Jianmin LI1()   

  1. 1.Department of Dermatology,Third Affiliated Hospital,Guangxi Medical University,Nanning 530031,China
    2.Department of Dermatology,First Affiliated Hospital,Guangxi Medical University,Nanning 530021,China
  • Received:2021-03-01 Online:2022-05-28 Published:2022-06-21
  • Contact: Jianmin LI E-mail:351615428@qq.com

Abstract: Objective

To investigate the inhibitory effect of silencing the expression of DNA methyltransferase 3A (DNMT3a) on the progression of psoriasis like HaCaT cell cycle and cell proliferation,and to clarify its possible mechanism.

Methods

The skin tissues of 15 patients with psoriasis vulgaris (psoriasis group) and 15 healthy controls (control group) were collected. Real-time fluorescence quantitative PCR(RT-qPCR) method was used to detect the expression levels of DNMT3a mRNA in skin tissue of the subjects in two groups. The Si-DNMT3a silenced by DNMT3a was transfected into the HaCaT cells by siRNA transfection technology. RT-qPCR and Western blotting methods were used to detect the transfection efficiency of si-DNMT3a. The HaCaT cells were induced by M5 method and the psoriasis like cell model was established. The cells were divided into control group, si-DNMT3a group, si-NC group, and HaCaT group. The proliferation rates of the cells in various groups were detected by CCK-8 method, the percentages of the cells at different cell cycles in various groups were detected by flow cytometry, and the apoptotic rates of the cells in various groups were detected by Annexin Ⅴ-FITC test. MSP test was used to detect the methylation levels of LATS1 gene promoter in skin tissue of the subjects in psoriasis group and control group. The expression levels of large tumor suppressor gene 1(LATS1), Yes associated protein(YAP1), cyclin D1, CDK6, and cleaved caspase-3 proteins in the HaCaT cells in various groups were detected by Western blotting method.

Results

Compared with control group, the expression level of DNMT3a mRNA in skin tissue of the patients in psoriasis group was significantly increased (P<0.01). Compared with HaCaT group,the expression levels of DNMT3a mRNA and protein in the HaCaT cells in si-NC group were decreased (P<0.05). Compared with HaCaT group, the proliferation rates of the HaCaT cells and the percentages of cells at S phase in control group, si -NC group,and si- DNMT3a group were increased (P<0.05), and the percentages of cells at G1 phase and the apoptotic rates were significantly decreased (P<0.05);compared with control group and si-NC group, the proliferation rates of the HaCaT cells and the percentage of cells at S phase in si-DNMT3a group were significantly decreased(P<0.05),and the apoptotic rate and the percentage of cells at G1 phase were increased (P<0.05). The MSP results showed that the methylation level of LATS1 promoter in skin tissue of the patients in psoriasis group was higher than that in control group (P<0.01). The Western blotting results showed that compared with HaCaT group, the expression levels of YAP1, Cyclin D1, and CDK6 proteins in the HaCaT cells in control group, si-NC group,and si-DNMT3a groups were increased (P<0.05), while the expression levels of LATS1 and cleaved caspase-3 proteins were decreased (P<0.05); compared with control group and si-NC group, the expression levels of YAP1, cyclin D1, and CDK6 proteins in the HaCaT cells in si- DNMT3a group were decreased (P<0.05), and the expression levels of LATS1 and cleaved caspase-3 proteins were increased (P<0.05).

Conclusion

Silencing the DNMT3a expression may down regulate the expression of YAP1 by reducing the methylation level of LATS1, so as to inhibit the progression of psoriasis like HaCaT cell cycle and cell proliferation and induce the apoptosis.

Key words: Psoriasis, DNA methyltransferases 3a, Large tumor suppressor gene 1, Yes-associated protein

CLC Number: 

  • R758.63