Mechanism of miR-214-5p in skin basal cell carcinoma through DNMT1-mediated DNA methylation modification of AXIN2 gene
Objective To investigate the effect of promoter methylation of axis inhibition protein 2(AXIN2)gene on gene transcription in cutaneous basal cell carcinoma(BCC)and the mechanismof miR-214-5p sregulation of Axis inhibition protein 2(AXIN2)methylation rate by targeting DNA methyltrans-ferase1(DNMT1).Methods From January 2022 to June 2023,102 BCC patients'sample and baseline da-ta who were treated in Guangzhou Institute of Dermatology were collected as the study objects.The meth-ylation rate of AXIN2 gene promoter region was quantitatively determined by pyrosequencing.real-time quantitative(PCR)was used to detect AXIN2,DNMT1 mRNA and miR-214-5p expression levels.The basal cell carcinoma A431 cells were transfected with miR-214-5p mimic,inhibitor and negative control mimic NC,respectively.The mRNA expression level of DNMT1 gene and the methylation rate of AXIN2 gene were detected 48 h later.Results The methylation rate of AXIN2 gene in BCC was significantly higher than that in paracanceral normal tissues(t=5.128,P＜0.001),and the mRNA relative expression level of AXIN2 gene was significantly lower than that in paracanceral normal tissues(t=7.826,P＜0.001).The mRNA expression level of DNMT1 gene was significantly higher than that of paracancerous normal tissues(t=4.838,P＜0.001),and the expression level of miR-214-5p was significantly lower than that of paracancerous normal tissues(t=5.426,P＜0.001).The methylation rate of AXIN2 gene in BCC cancer tissue was negatively correlated with its mRNA expression level(r=-0.793,P＜0.001),and the mRNA level of DNMT1 gene was positively correlated with the methylation rate of AXIN2 gene(r=0.814,P＜0.001).The expression level of miR-214-5p was negatively correlated with the mRNA level of DNMT1 gene(r=-0.747,P＜0.001).The results of double luciferase reporter gene experiment con-firmed that DNMT1 was the target gene of miR-214-5p.After transfection,the mRNA levels of DNMT1 gene and the methylation rate of AXIN2 gene in mimic were significantly decreased compared with those of mimic NC,inhibitor and inhibitor NC(P＜0.001).The mRNA level of DNMT1 gene and the methyla-tion rate of AXIN2 gene in inhibitor were significantly increased compared with the other three(P＜0.001).Conclusion miR-214-5p can affect the DNA methylation rate of AXIN2 gene by regulating the ex-pression of downstream target protein DNMT1,regulate the expression level of AXIN2 gene and partici-pate in the pathogenesis of skin basal cell carcinoma.
basal cell carcinomamiR-214-5pDNA methyltransferase1axial inhibitory protein2promot-er region methylation
万方数据
维普
