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首页> 外文期刊>Cell biology international. >Abnormal expression of DNA methyltransferases and genomic imprinting in cloned goat fibroblasts
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Abnormal expression of DNA methyltransferases and genomic imprinting in cloned goat fibroblasts

机译：克隆山羊成纤维细胞中DNA甲基转移酶的异常表达和基因组印迹

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摘要

Somatic cell nuclear transfer (SCNT) is a useful way to produce cloned animals. However, SCNT animals exhibit DNA methylation and genomic imprinting abnormalities. These abnormalities may be due to the faulty epigenetic reprogramming of donor cells. To investigate the consequence of SCNT on the genomic imprinting and global methylation in the donor cells, growth patterns and apoptosis of cloned goat fibroblast cells (CGFCs) at passage 7 were determined. Growth patterns in CGFCs were similar to the controls; however, the growth rate in log phase was lower and apoptosis in CGFCs were significantly higher (P < 0.01). In addition, quantitative expression analysis of three DNA methyltransferases (Dnmt) and two imprinted genes (H19, IGF2R) was conducted in CGFCs: Dnmt1 and Dnmt3b expression was significantly reduced (P < 0.01), and H19 expression was decreased sixfold (P < 0.01); however, the expression of Dnmt3a was unaltered and IGF2R expression was significantly increased (P < 0.05). Finally, we used bisulfite sequencing PCR to compare the DNA methylation patterns in differentially methylated regions (DMRs) of H19 and IGF2R. The DMRs of H19 (P < 0.01) and IGF2R (P < 0.01) were both highly methylated in CGFCs. These results indicate that the global genome might be hypomethylated. Moreover, there is an aberrant expression of imprinted genes and DMR methylation in CGFCs.
机译：体细胞核移植（SCNT）是生产克隆动物的有用方法。但是，SCNT动物表现出DNA甲基化和基因组印迹异常。这些异常可能是由于供体细胞的表观遗传重编程错误。为了研究SCNT对供体细胞中基因组印迹和整体甲基化的影响，确定了第7代克隆山羊成纤维细胞（CGFC）的生长模式和凋亡。 CGFCs的生长方式与对照相似。然而，对数期的生长速率较低，CGFCs的细胞凋亡明显较高（P <0.01）。此外，在CGFC中对三种DNA甲基转移酶（Dnmt）和两个印迹基因（H19，IGF2R）进行了定量表达分析：Dnmt1和Dnmt3b的表达显着降低（P <0.01），H19的表达降低了六倍（P <0.01）。 ）;然而，Dnmt3a的表达未改变，而IGF2R的表达则显着增加（P <0.05）。最后，我们使用亚硫酸氢盐测序PCR来比较H19和IGF2R的差异甲基化区域（DMR）中的DNA甲基化模式。 H19（P <0.01）和IGF2R（P <0.01）的DMR在CGFC中均高度甲基化。这些结果表明，全局基因组可能被低甲基化。此外，CGFC中的印迹基因和DMR甲基化异常表达。

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