Mechanism of UPF1 Methylation and miR-744-5p/CCND1 in Papillary Thyroid Carcinoma
Objective:To investigate the mechanism of UPF1 methylation and miR-744-5p/CCND1 in papillary thyroid cancer.Methods:The human papillary thyroid cancer cell line TCP-1 and normal thyroid epithelial cell Nthy ori-3 were intervened with demethylation reagent 5-Aza CdR,respectively.Methylation specific PCR technology was used to detect the methylation changes of UPF1 gene before and after the intervention,and Western blotting was used to detect the relative expression of intervention UPF1,DN-MT1,miR-744-5p,and CCND1 proteins.Results:PCR amplification showed that the UPF1 gene only showed non methylated primer amplification bands(U bands)in the Nthy ori-3 group,and only methylated primer amplification bands(M bands)in the TCP-1 group.After the action of 5-Aza-Cdr,the methylation amplification bands of UPF1 gene decreased,and the methylation expression decreased.There was a statistically significant difference in the relative expression of UPF1,DNMT1,miR-744-5p,and CCND1 proteins among each group(P＜0.05).Compared with the Nthy ori-3 group,the relative expression of DNMT1 and UPF1 proteins in the TCP-1 group was significantly increased,while the relative expression of miR-744-5p and CCND1 proteins was significantly reduced(P＜0.05);Compared with the TCP-1 group,the relative expression of DNMT1 and UPF1 proteins in the TCP-1 intervention group was significantly reduced,while the relative expression of miR-744-5p and CCND1 proteins was significantly increased(P＜0.05).Compared with the TCP-1 group,the number of cell invasion and migration in the TCP-1 intervention group was significantly reduced(P＜0.05).Conclusion:UPF1 methy-lation is present in papillary thyroid cancer,and the expression loss of UPF1 gene methylation may inhibit the miR-744-5p/CCND1 axis,playing a key role in the occurrence and development of papillary thyroid cancer.
万方数据
维普
