High‐risk HPVs (HR‐HPVs) are DNA viruses considered as primary etiologic factors in malignancies of the low female genital tract. Their presence has also been documented in oropharyngeal and laryngeal cancers. However, HPV infection is considered a necessary but not sufficient cause of tumoral development; meantime, increasing evidences on the tumorigenic role of cancer stem cells (CSCs) have been documented in the literature. CSCs represent a small subpopulation of neoplastic cells with self‐renewal potential, capable of maintaining tumor growth and cell differentiation, also involved in metastatic process, recurrence, and resistance to chemotherapeutic agents. In the present study, performed on KB cell lines, we evaluated the tumor forming potential of CSCs, and their relationship with the HPV infection status. We started our study by identifying the most aggressive cell line on the minimal number of cells being able of growth in vivo in a model of athymic nude mice (BALB/c nu/nu). We used an oral‐derived KB cell line separated in the KB‐CD133+ and KB‐CD133‐ populations, by using immunomagnetic beads and fluorescence‐activated cell sorting (FACS). The separated populations were injected in athymic nude mice (BALB/c nu/nu). Xenograft tumors have been analyzed for tumor size, CD133 expression by immunohistochemistry (IHC) and for DNA HR‐HPV integration by in situ hybridization (ISH), comparing CD133‐enriched xenograft tumors versus the CD133 non‐enriched ones. On standard conditions, the KB cell line has a poor population of glycosylated CD133 marker (<5.0%) when investigated with antibodies versus CD133, and more specifically its glycosylated epitope (AC133). Enriched CD133 KB cells possess a higher capacity of tumor growth in xenograft models of nude mice when compared to KB CD133‐negative cells. We observed that the AC133 epitope, extensively used to purifying hematopoietic stem cells, is able to select an epithelial subpopulation of cancer stem cells with aggressive behavior. We retain that CD133 may be a useful target in anticancer strategies including pharmacological and immunological therapies.

中文翻译：

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HR-HPV与干细胞在肿瘤形成过程中的可能相互作用：CD133作为HPV18感染的kb细胞系中癌症干细胞的推定标记的体内研究。

高危HPV（HR-HPV）是DNA病毒，被认为是女性低生殖道恶性肿瘤的主要病因。在口咽癌和喉癌中也已发现它们的存在。但是，HPV感染被认为是肿瘤发展的必要但非充分的原因。同时，文献中已经记录了关于癌干细胞（CSCs）致癌作用的越来越多的证据。CSC代表具有自我更新潜能的一小部分肿瘤细胞，能够维持肿瘤生长和细胞分化，还参与转移过程，复发和对化学治疗药物的耐药性。在本研究中，我们对KB细胞系进行了评估，评估了CSCs的肿瘤形成潜力，以及它们与HPV感染状态的关系。我们通过在无胸腺裸鼠（BALB / c nu / nu）模型中能够在体内最小生长的​​细胞中鉴定出最具攻击性的细胞系来开始我们的研究。我们通过免疫磁珠和荧光激活细胞分选（FACS），使用了在KB‐CD133 +和KB‐CD133‐人群中分离的口腔来源KB细胞系。将分离的群体注射到无胸腺裸鼠（BALB / c nu / nu）中。已通过免疫组织化学（IHC）分析了异种移植瘤的肿瘤大小，CD133表达，并通过原位杂交（ISH）分析了DNA HR-HPV整合，比较了富含CD133的异种移植瘤与未富含CD133的异种移植瘤。在标准条件下，用抗体对CD133进行研究时，KB细胞系的糖基化CD133标记物种群较少（<5.0％），更具体地讲，其糖基化表位（AC133）。与KB CD133阴性细胞相比，在裸鼠异种移植模型中，富含CD133 KB细胞具有更高的肿瘤生长能力。我们观察到，广泛用于纯化造血干细胞的AC133表位能够选择具有攻击行为的癌症干细胞的上皮亚群。我们保留CD133可能是抗癌策略（包括药物和免疫疗法）中的有用靶标。