[丝裂霉素C处理HeLa细胞miRNA异常表达对DNA甲基化的依赖性]。

Genetika Pub Date : 2016-11-01
V A Tarasov, N V Boyko, M A Makhotkin, E F Shin, M G Tyutyakina, I E Chikunov, A V Naboka, A N Mashkarina, A A Kirpiy, D G Matishov
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引用次数: 0

摘要

研究丝裂霉素C处理8天后HeLa细胞中miRNAs及其前体(pre-miRNAs)的表达对DNA甲基化水平的依赖性。采用大规模平行DNA测序法分析miRNA表达。5-氮杂胞苷(DNA甲基化抑制剂)在诱变剂暴露6天后加入培养基。结果表明,一些成熟mirna(61个中的39个)的表达变化伴随着它们的pre-miRNA的表达变化,而其他成熟mirna(61个中的22个)的pre-miRNA的表达没有明显变化。5-氮胞苷处理后,61个成熟mirna中的8个和55个预mirna中的6个在诱导的HeLa细胞中保持了异常表达。此外,在没有丝裂霉素C处理的HeLa细胞中,超过90%的mirna的表达不依赖或仅依赖于DNA甲基化水平,这表明丝裂霉素C处理后表达发生了显著变化。结果表明丝裂霉素C诱导异常DNA甲基化,影响突变原处理后细胞世代miRNA表达变化的维持。
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[The miRNA aberrant expression dependence on DNA methylation in HeLa cells treated with mitomycin C].

The dependence of expression of miRNAs and their precursors (pre-miRNAs) on the DNA methylation level in HeLa cells 8 days after mitomycin C treatment was studied. A massive parallel DNA sequencing method was applied to analyze miRNA expression. 5-Azacytidine (DNA methylation inhibitor) was added to the medium 6 days after mutagenic agent exposure. The results indicated that the change in expression for some mature miRNAs (39 of 61) was accompanied by the change in the expression of their pre-miRNAs, while there were no significant changes in the expression of pre-miRNA for other mature miRNAs (22 of 61). The aberrant expression was maintained by 8 of 61 mature miRNAs and 6 of 55 pre-miRNAs in the induced HeLa cells after 5-azacytidine treatment. In addition, the expression of more than 90% of miRNAs, which indicated a significant change in expression after mitomycin C treatment, does not depend or depends slightly on the DNA methylation level in HeLa cells without mitomycin C treatment. The results suggest that mitomycin C induces aberrant DNA methylation which affects maintenance of changes in the miRNA expression in cell generations after mutagen treatment.

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