Institute of Virology, Vetsuisse Faculty, University of Zurich, 8057 Zurich, Switzerland.
Dermatology Unit, Vetsuisse Faculty, University of Zurich, 8057 Zurich, Switzerland.
Viruses. 2021 Jul 19;13(7):1404. doi: 10.3390/v13071404.
There is growing evidence that equine papillomavirus type 2 (EcPV2) infection is etiologically associated with the development of genital squamous cell carcinoma (SCC) and precursor lesions in equids. However, the precise mechanisms underlying neoplastic progression remain unknown. To allow the study of EcPV2-induced carcinogenesis, we aimed to establish a primary equine cell culture model of EcPV2 infection. Three-dimensional (3D) raft cultures were generated from equine penile perilesional skin, plaques and SCCs. Using histological, molecular biological and immunohistochemical methods, rafts versus corresponding natural tissue sections were compared with regard to morphology, presence of EcPV2 DNA, presence and location of EcPV2 gene transcripts and expression of epithelial, mesenchymal and tumor/proliferation markers. Raft cultures from perilesional skin harboring only a few EcPV2-positive (EcPV2+) cells accurately recapitulated the differentiation process of normal skin, whilst rafts from EcPV2+ penile plaques were structurally organized but showed early hyperplasia. Rafts from EcPV2+ SCCs exhibited pronounced hyperplasia and marked dysplasia. Raft levels of EcPV2 oncogene transcription (E6/E7) and expression of tumor/proliferation markers p53, Ki67 and MCM7 expression positively correlated with neoplastic progression, again reflecting the natural situation. Three-dimensional raft cultures accurately reflected major features of corresponding ex vivo material, thus constituting a valuable new research model to study EcPV2-induced carcinogenesis.
越来越多的证据表明，马乳头瘤病毒 2 型（EcPV2）感染与马属动物的生殖器鳞状细胞癌（SCC）和前体病变的发生有关。然而，肿瘤进展的确切机制仍不清楚。为了研究 EcPV2 诱导的致癌作用，我们旨在建立 EcPV2 感染的马原代细胞培养模型。从马阴茎病变周围皮肤、斑块和 SCC 中生成了三维（3D）筏培养物。使用组织学、分子生物学和免疫组织化学方法，将筏培养物与相应的天然组织切片进行比较，比较了形态、EcPV2 DNA 的存在、EcPV2 基因转录本的存在和位置以及上皮、间充质和肿瘤/增殖标志物的表达。仅含有少数 EcPV2 阳性（EcPV2+）细胞的病变周围皮肤筏培养物准确地再现了正常皮肤的分化过程，而来自 EcPV2+阴茎斑块的筏培养物结构上是有组织的，但表现出早期增生。来自 EcPV2+SCC 的筏培养物表现出明显的增生和显著的发育不良。筏培养物中 EcPV2 致癌基因转录（E6/E7）和肿瘤/增殖标志物 p53、Ki67 和 MCM7 的表达与肿瘤进展呈正相关，再次反映了自然情况。三维筏培养物准确地反映了相应的离体材料的主要特征，因此构成了研究 EcPV2 诱导的致癌作用的有价值的新研究模型。
