INTRODUCTION The main cause of cervical cancer is an infection of keratinocytes in the basal layer of the stratified epithelium of the cervix by human papillomavirus (HPV). Other than in cervical samples, HPV DNA has been found in serum and other fluids but its origin is unclear. Extracellular vesicles (EV) could be a conveyance of viral DNA given their emerging role in cellular communication. The content of EV derived from cervical cells has not been properly explored and it is not known whether or not they contain HPV DNA. METHODS We evaluated the DNA content of exosomes purified from cultures of HeLa cells by Next Generation Sequencing (NGS) and confirmed its presence by PCR. The presence of HPV DNA was also evaluated by PCR and NGS in EV from HPV-positive cervical samples without apparent lesion or with LSIL. RESULTS We detected the integrated form of viral-DNA in exosomes from HeLa cells by NGS and confirmed its presence by PCR. The search for HPV sequences in EV obtained from cervical exudate samples without apparent lesion or with LSIL, where we expected to find the viral genome as an episome, indicated that HPV DNA, including the E6 and E7 oncogenes, is present in these EV. CONCLUSION HPV DNA, including the viral oncogenes E6/E7, is found in exosomes regardless of the integration status of the virus in the infected cell.

中文翻译：

---

来自 HeLa 细胞和宫颈样本的细胞外囊泡中存在 HPV DNA。

引言 宫颈癌的主要原因是宫颈复层上皮基底层角质形成细胞被人乳头瘤病毒 (HPV) 感染。除宫颈样本外，还在血清和其他体液中发现了 HPV DNA，但其来源尚不清楚。鉴于它们在细胞通讯中的新兴作用，细胞外囊泡 (EV) 可能是病毒 DNA 的载体。来自宫颈细胞的 EV 的含量尚未得到适当探索，也不知道它们是否含有 HPV DNA。方法我们通过下一代测序 (NGS) 评估从 HeLa 细胞培养物中纯化的外泌体的 DNA 含量，并通过 PCR 确认其存在。HPV DNA 的存在也通过 PCR 和 NGS 在来自无明显病变或 LSIL 的 HPV 阳性宫颈样本的 EV 中进行评估。结果 我们通过 NGS 检测到 HeLa 细胞外泌体中病毒 DNA 的整合形式，并通过 PCR 确认其存在。从没有明显病变或具有 LSIL 的宫颈渗出液样本中寻找 EV 中的 HPV 序列，我们希望在其中找到病毒基因组作为附加体，表明 HPV DNA，包括 E6 和 E7 癌基因，存在于这些 EV 中。结论 无论病毒在受感染细胞中的整合状态如何，都在外泌体中发现了 HPV DNA，包括病毒癌基因 E6/E7。存在于这些 EV 中。结论 无论病毒在感染细胞中的整合状态如何，都在外泌体中发现了 HPV DNA，包括病毒癌基因 E6/E7。存在于这些 EV 中。结论 无论病毒在感染细胞中的整合状态如何，都在外泌体中发现了 HPV DNA，包括病毒癌基因 E6/E7。