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Abstract：
Previous studies have shown that the BMP7 gene is differentially expressed in Hu sheep lamb skin of different pattern types, and its expression level is significantly correlated with hair follicle indices of different pattern types, but the molecular mechanism of the differential expression of the BMP7 gene remains unclear. This study investigated the effect of DNA methylation on the transcriptional expression of BMP7. Firstly, we found that the mRNA expression of the BMP7 gene and the activity of the core promoter of the BMP7 gene were upregulated after 5-Aza-Deoxycytidine-induced demethylation treatment using qRT-PCR and double luciferase reporter assay. Then, we found that the proliferation of Hu sheep DPCs in vitro was promoted after 5-Aza-Deoxycytidine-induced demethylation treatment through qRT-PCR, CCK-8, and EdU assay, and that the overexpression of DNMT1 in DPCs induced the opposite effect. In addition, the results of the cell cycle assay reveal that the percentage of cells in the S phase was increased after 5-Aza-Deoxycytidine-induced demethylation treatment, and that the percentage of cells in the S phase was decreased after overexpression of DNMT1 in DPCs. This study indicated that the differential expression of the BMP7 gene in different patterns of Hu sheep lamb skin may be regulated by DNA methylation modification. In addition, DNA methylation can regulate the proliferation and cell cycle of DPCs in Hu sheep.
摘要：
前期研究表明,BMP7基因在不同形态类型的湖羊羔羊皮中差异表达,其表达水平与不同类型毛囊指数显著相关,但BMP7基因差异表达的分子机制尚不清楚。本研究探讨了DNA甲基化对BMP7转录表达的影响。首先,我们发现,使用qRT-PCR和双荧光素酶报告基因分析,在5-Aza-脱氧胞苷诱导的去甲基化处理后,BMP7基因的mRNA表达和BMP7基因核心启动子的活性上调.然后，我们发现，通过qRT-PCR，5-氮杂脱氧胞苷诱导的去甲基化处理后，湖羊DPCs的体外增殖得到促进，CCK-8和EdU分析，DNMT1在DPC中的过表达诱导了相反的作用。此外,细胞周期测定结果表明，在5-氮杂脱氧胞苷诱导的去甲基化处理后，S期细胞的百分比增加，DNMT1在DPC中过表达后，S期细胞的百分比降低。本研究表明，BMP7基因在不同模式湖羊羊皮中的差异表达可能受到DNA甲基化修饰的调控。此外,DNA甲基化可以调控湖羊DPCs的增殖和细胞周期。
