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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Chromosomal insertion of foreign (adenovirus type 12, plasmid, or bacteriophage lambda) DNA is associated with enhanced methylation of cellular DNA segments.
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Chromosomal insertion of foreign (adenovirus type 12, plasmid, or bacteriophage lambda) DNA is associated with enhanced methylation of cellular DNA segments.

机译：外源（12型腺病毒，质粒或噬菌体λ）DNA的染色体插入与细胞DNA片段甲基化增强有关。

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摘要

Insertion of foreign DNA into an established mammalian genome can extensively alter the patterns of cellular DNA methylation. Adenovirus type 12 (Ad12)-transformed hamster cells, Ad12-induced hamster tumor cells, or hamster cells carrying integrated DNA of bacteriophage lambda were used as model systems. DNA methylation levels were examined by cleaving cellular DNA with Hpa II, Msp I, or Hha I, followed by Southern blot hybridization with 32P-labeled, randomly selected cellular DNA probes. For several, but not all, cellular DNA segments investigated, extensive increases in DNA methylation were found in comparison with the methylation patterns in BHK21 or primary Syrian hamster cells. In eight different Ad12-induced hamster tumors, moderate increases in DNA methylation were seen. Increased methylation of cellular genes was also documented in two hamster cell lines with integrated Ad12 DNA without the Ad12-transformed phenotype, in one cloned BHK21 cell line with integrated plasmid DNA, and in at least three cloned BHK21 cell lines with integrated lambda DNA. By fluorescent in situ hybridization, the cellular hybridization probes were located to different hamster chromosomes. The endogenous intracisternal A particle genomes showed a striking distribution on many hamster chromosomes, frequently on their short arms. When BHK21 hamster cells were abortively infected with Ad12, increases in cellular DNA methylation were not seen. Thus, Ad12 early gene products were not directly involved in increasing cellular DNA methylation. We attribute the alterations in cellular DNA methylation, at least in part, to the insertion of foreign DNA. Can alterations in the methylation profiles of hamster cellular DNA contribute to the generation of the oncogenic phenotype?
机译：将外源DNA插入已建立的哺乳动物基因组中可以大大改变细胞DNA甲基化的模式。使用腺病毒12型（Ad12）转化的仓鼠细胞，Ad12诱导的仓鼠肿瘤细胞或携带整合有噬菌体λDNA的仓鼠细胞作为模型系统。通过用Hpa II，Msp I或Hha I裂解细胞DNA，然后与32P标记的随机选择的细胞DNA探针进行Southern杂交，检查DNA甲基化水平。对于一些但不是全部的细胞DNA片段，与BHK21或原代叙利亚仓鼠细胞的甲基化模式相比，发现了DNA甲基化的广泛增加。在8种不同的Ad12诱导的仓鼠肿瘤中，观察到DNA甲基化的适度增加。在两个整合了Ad12 DNA而没有Ad12转化表型的仓鼠细胞系中，在一个克隆了整合质粒DNA的BHK21细胞系中，以及至少三个克隆了Lambda DNA整合的BHK21细胞系中，也记录了细胞基因甲基化的增加。通过荧光原位杂交，将细胞杂交探针定位于不同的仓鼠染色体。内源性脑内壳内A颗粒基因组在许多仓鼠染色体上表现出惊人的分布，通常在它们的短臂上。当BHK21仓鼠细胞被Ad12流产感染时，未见细胞DNA甲基化增加。因此，Ad12早期基因产物并不直接参与增加细胞DNA甲基化。我们将细胞DNA甲基化的变化至少部分归因于外源DNA的插入。仓鼠细胞DNA甲基化谱的改变是否有助于致癌表型的产生？

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