Department of Otorhinolaryngology Head and Neck Surgery, University Hospital Mannheim, University of Heidelberg, 68167 Mannheim, Germany.
Department of Pathology, University of Pittsburgh School of Medicine and University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213, USA.
Int J Mol Sci. 2020 Nov 13;21(22):8570. doi: 10.3390/ijms21228570.
Human papillomavirus (HPV)(+) and HPV(-) head and neck cancer (HNC) cells' interactions with the host immune system are poorly understood. Recently, we identified molecular and functional differences in exosomes produced by HPV(+) vs. HPV(-) cells, suggesting that genetic cargos of exosomes might identify novel biomarkers in HPV-related HNCs. Exosomes were isolated by size exclusion chromatography from supernatants of three HPV(+) and two HPV(-) HNC cell lines. Paired cell lysates and exosomes were analyzed for messenger RNA (mRNA) by qRT-PCR and microRNA (miR) contents by nanostring analysis. The mRNA profiles of HPV(+) vs. HPV(-) cells were distinct, with , and overexpressed in HPV(+) cells and , and in HPV(-) cells. The mRNA profiles of HPV(+) or HPV(-) exosomes resembled the cargo of their parent cells. miR expression profiles in cell lysates identified 8 miRs expressed in HPV(-) cells vs. 14 miRs in HPV(+) cells. miR-205-5p was exclusively expressed in HPV(+) exosomes, and miR-1972 was only detected in HPV(-) exosomes. We showed that HPV(+) and HPV(-) exosomes recapitulated the mRNA expression profiles of their parent cells. Expression of miRs was dependent on the HPV status, and miR-205-5p in HPV(+) and miR-1972 in HPV(-) exosomes emerge as potential discriminating HPV-associated biomarkers.
人乳头瘤病毒（HPV）（+）和 HPV（-）头颈部癌症（HNC）细胞与宿主免疫系统的相互作用知之甚少。最近，我们发现 HPV（+）与 HPV（-）细胞产生的外泌体在分子和功能上存在差异，这表明外泌体的遗传 cargos 可能鉴定出 HPV 相关 HNC 的新型生物标志物。外泌体通过大小排阻色谱法从三种 HPV（+）和两种 HPV（-）HNC 细胞系的上清液中分离得到。对配对的细胞裂解物和外泌体进行信使 RNA（mRNA）的 qRT-PCR 分析和 microRNA（miR）含量的纳米串分析。HPV（+）与 HPV（-）细胞的 mRNA 谱明显不同，HPV（+）细胞中过度表达 、 和 ，而 HPV（-）细胞中则过度表达 、 和 。HPV（+）或 HPV（-）外泌体的 mRNA 谱与其亲本细胞的货物相似。细胞裂解物中的 miR 表达谱鉴定出 8 种在 HPV（-）细胞中表达的 miR 和 14 种在 HPV（+）细胞中表达的 miR。miR-205-5p 仅在 HPV（+）外泌体中表达，而 miR-1972 仅在 HPV（-）外泌体中检测到。我们表明，HPV（+）和 HPV（-）外泌体再现了其亲本细胞的 mRNA 表达谱。miR 的表达依赖于 HPV 的状态，HPV（+）中的 miR-205-5p 和 HPV（-）中的 miR-1972 在外泌体中作为潜在的区分 HPV 相关生物标志物出现。
