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首页> 外文期刊>International Journal of Cancer =: Journal International du Cancer >Identification of human papillomavirus (HPV) 16 DNA integration and the ensuing patterns of methylation in HPV-associated head and neck squamous cell carcinoma cell lines
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Identification of human papillomavirus (HPV) 16 DNA integration and the ensuing patterns of methylation in HPV-associated head and neck squamous cell carcinoma cell lines

机译：鉴定人乳头瘤病毒（HPV）16 DNA集成以及HPV相关头颈鳞状细胞癌细胞系中的甲基化模式

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摘要

Recent studies showed that human papillomavirus (HPV) integration contributes to the genomic instability seen in HPV-associated head and neck squamous cell carcinoma (HPV-HNSCC). However, the epigenetic alterations induced after HPV integration remains unclear. To identify the molecular details of HPV16 DNA integration and the ensuing patterns of methylation in HNSCC, we performed next-generation sequencing using a target-enrichment method for the effective identification of HPV16 integration breakpoints as well as the characterization of genomic sequences adjacent to HPV16 integration breakpoints with three HPV16-related HNSCC cell lines. The DNA methylation levels of the integrated HPV16 genome and that of the adjacent human genome were also analyzed by bisulfite pyrosequencing. We found various integration loci, including novel integration sites. Integration loci were located predominantly in the intergenic region, with a significant enrichment of the microhomologous sequences between the human and HPV16 genomes at the integration breakpoints. Furthermore, various levels of methylation within both the human genome and the integrated HPV genome at the integration breakpoints in each integrant were observed. Allele-specific methylation analysis suggested that the HPV16 integrants remained hypomethylated when the flanking host genome was hypomethylated. After integration into highly methylated human genome regions, however, the HPV16 DNA became methylated. In conclusion, we found novel integration sites and methylation patterns in HPV-HNSCC using our unique method. These findings may provide insights into understanding of viral integration mechanism and virus-associated carcinogenesis of HPV-HNSCC.
机译：最近的研究表明，人乳头瘤病毒（HPV）一体化有助于HPV相关头颈鳞状细胞癌（HPV-HWNSCC）中所见的基因组不稳定性。然而，HPV集成后诱导的表观遗传改变仍然不清楚。为了鉴定HPV16 DNA积分的分子细节和HPNSCC中的甲基化模式，我们使用靶向鉴定HPV16集成断点的有效鉴定的下一代测序以及与HPV16集成相邻的基因组序列的表征具有三种HPV16相关的HNSCC细胞系的断点。通过双硫酸氢盐焦肌肉，还分析了集成HPV16基因组的DNA甲基化水平和相邻的人类基因组的水平。我们发现各种集成基因座，包括新颖的集成站点。积分基因座主要位于非基因区域，在整合断点处的人和HPV16基因组之间的微甾级序列具有显着性富集。此外，观察到每种整合体中的整体断裂点的人类基因组和集成的HPV基因组中的各种甲基化水平。等位基因特异性甲基化分析表明，当侧翼宿主基因组是脱甲酰化时，HPV16整合体仍保持着黄色甲基化。然而，在整合到高度甲基化的人类基因组区域之后，HPV16 DNA变为甲基化。总之，我们使用我们独特的方法在HPV-HNSCC中发现了新的集成位点和甲基化模式。这些发现可以提供对HPV-HNSCC的病毒整合机制和病毒相关致癌物理的见解。

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