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.
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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DOI：
10.1002/ijc.30589
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发表时间：
2017-04-01
影响因子：
6.4
通讯作者：
Oridate N
中科院分区：
文献类型：
--
作者：
Hatano T;Sano D;Takahashi H;Hyakusoku H;Isono Y;Shimada S;Sawakuma K;Takada K;Oikawa R;Watanabe Y;Yamamoto H;Itoh F;Myers JN;Oridate N
最近的研究表明，人乳头瘤病毒（HPV）的整合有助于在HPV相关的头颈部鳞状细胞癌（HPV-HNSCC）中观察到的基因组不稳定性。然而，HPV整合后诱导的表观遗传学改变仍不清楚。为了确定HPV 16 DNA整合的分子细节和HNSCC中随后的甲基化模式，我们使用靶富集方法进行了下一代测序，以有效鉴定HPV 16整合断点，并对三种HPV 16相关HNSCC细胞系的HPV 16整合断点附近的基因组序列进行表征。通过亚硫酸氢盐焦磷酸测序分析整合的HPV 16基因组和相邻人类基因组的DNA甲基化水平。我们发现了各种整合位点，包括新的整合位点。整合位点主要位于基因间区域，在整合断点处人和HPV 16基因组之间的微同源序列显著富集。此外，在每个整合体的整合断点处，观察到人类基因组和整合的HPV基因组内的不同水平的甲基化。等位基因特异性甲基化分析表明，当侧翼宿主基因组低甲基化时，HPV 16整合体保持低甲基化。然而，在整合到高度甲基化的人类基因组区域后，HPV 16 DNA变得甲基化。总之，我们使用我们独特的方法在HPV-HNSCC中发现了新的整合位点和甲基化模式。这些发现可能为理解HPV-HNSCC的病毒整合机制和病毒相关的致癌作用提供了新的见解。
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.
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影响因子：
6.2
作者：
Reuschenbach, Miriam;Huebbers, Christian U.;Doeberitz, Magnus von Knebel
通讯作者：
Doeberitz, Magnus von Knebel
影响因子：
8
作者：
Thorland, EC;Myers, SL;Smith, DI
通讯作者：
Smith, DI
影响因子：
6.4
作者：
Chaiwongkot, Arkom;Vinokurova, Svetlana;Doeberitz, Magnus von Knebel
通讯作者：
Doeberitz, Magnus von Knebel
影响因子：
--
作者：
Liu Y;Lu Z;Xu R;Ke Y
通讯作者：
Ke Y
影响因子：
3.3
作者：
Sano, Daisuke;Oridate, Nobuhiko
通讯作者：
Oridate, Nobuhiko