Human papillomavirus-related oropharyngeal squamous cell carcinoma (HPV+ OPSCC) represents a unique disease entity within head and neck cancer with rising incidence. Previous work has shown that alternative splicing events (ASEs) are prevalent in HPV+ OPSCC, but further validation is needed to understand the regulation of this process and its role in these tumours. In this study, eleven ASEs (GIT2, CTNNB1, MKNK2, MRPL33, SIPA1L3, SNHG6, SYCP2, TPRG1, ZHX2, ZNF331, and ELOVL1) were selected for validation from 109 previously published candidate ASEs to elucidate the post-transcriptional mechanisms of oncogenesis in HPV+ disease. In vitro qRT-PCR confirmed differential expression of 9 of 11 ASE candidates, and in silico analysis within the TCGA cohort confirmed 8 of 11 candidates. Six ASEs (MRPL33, SIPA1L3, SNHG6, TPRG1, ZHX2, and ELOVL1) showed significant differential expression across both methods. Further evaluation of chromatin modification revealed that ASEs strongly correlated with cancer-specific distribution of acetylated lysine 27 of histone 3 (H3K27ac). Subsequent epigenetic treatment of HPV+ HNSCC cell lines (UM-SCC-047 and UPCI-SCC-090) with JQ1 not only induced downregulation of cancer-specific ASE isoforms, but also growth inhibition in both cell lines. The UPCI-SCC-090 cell line, with greater ASE expression, also showed more significant growth inhibition after JQ1 treatment. This study confirms several novel cancer-specific ASEs in HPV+OPSCC and provides evidence for the role of chromatin modifications in regulation of alternative splicing in HPV+OPSCC. This highlights the role of epigenetic changes in the oncogenesis of HPV+OPSCC, which represents a unique, unexplored target for therapeutics that can alter the global post-transcriptional landscape.

人乳头瘤病毒相关的口咽鳞状细胞癌（HPV + OPSCC）代表着头颈癌中一种独特的疾病实体，且发病率不断上升。先前的工作表明，替代剪接事件（ASE）在HPV + OPSCC中很普遍，但是需要进一步验证才能了解该过程的调控及其在这些肿瘤中的作用。在这项研究中，从109个先前发表的候选ASE中选择了11种ASE（GIT2，CTNNB1，MKNK2，MRPL33，SIPA1L3，SNHG6，SYCP2，TPRG1，ZHX2，ZNF331和ELOVL1）进行验证，以阐明肿瘤发生后转录机制HPV +疾病。体外qRT-PCR证实了11种ASE候选药物中的9种在计算机中的差异表达TCGA队列中的分析确认了11名候选人中的8名。六个ASE（MRPL33，SIPA1L3，SNHG6，TPRG1，ZHX2和ELOVL1）两种方法均显示出显着的差异表达 染色质修饰的进一步评估表明，ASE与组蛋白3（H3K27ac）的乙酰化赖氨酸27的癌症特异性分布密切相关。随后用JQ1对HPV + HNSCC细胞系（UM-SCC-047和UPCI-SCC-090）进行表观遗传处理，不仅诱导了癌特异性ASE亚型的下调，而且还抑制了这两种细胞系的生长。具有更高的ASE表达的UPCI-SCC-090细胞系在JQ1处理后也显示出更显着的生长抑制作用。这项研究证实了HPV + OPSCC中几种新颖的癌症特异性ASE，并提供了染色质修饰在HPV + OPSCC中选择性剪接调控中的作用的证据。这突显了表观遗传学变化在HPV + OPSCC肿瘤发生中的作用，代表了独特的，