Introduction: Persistent high-risk human papillomavirus (hr-HPV) infection is the primary cause of cervical cancer. Integration of the HPV genome into the host chromosome is a key carcinogenic event. However, previous research has focused on alterations in genes adjacent to integration sites, leaving long-range effects on distal genes poorly understood. This study aimed to investigate how HPV integration at 13q22.1 distally regulates expression of diacylglycerol kinase θ (DGKH) and elucidate DGKH's biological functions and molecular mechanisms in cervical carcinogenesis.
Methods: We performed bioinformatics analysis using multi-omics data from public databases and our cohort to identify distally upregulated genes by comparing transcriptomes of patients with or without HPV integration at 13q22.1. Functional validation was conducted using a 13q22.1-specific HPV knock-in HaCaT cell model and cervical cancer cell lines through Western blot, siRNA interference, plasmid transfection, CCK-8, and Transwell assays. Mechanistic insights were explored using RNA interference, ChIP-PCR, and RNA sequencing.
Results: DGKH was upregulated in cervical cancer samples with HPV integration at 13q22.1. Its high expression correlated with poor patient prognosis and low CD8+ T cell infiltration. Knockdown of DGKH inhibited, while overexpression promoted, proliferation, migration, and invasion of cervical cancer cells. The transcription factor ELF1, upregulated post-integration, directly binds to the DGKH promoter to activate transcription. Furthermore, DGKH enhanced macroautophagy, which may facilitate tumor cell survival under stress.
Conclusions: This study demonstrates that HPV integration can exert long-range transcriptional regulation on distal genes. We reveal a critical oncogenic role of DGKH in cervical cancer, where its expression is upregulated via ELF1 following HPV integration. DGKH promotes tumor proliferation and invasion, likely by enhancing macroautophagy, leading to poor prognosis. These findings deepen understanding of HPV-driven carcinogenesis and identify DGKH as a potential prognostic biomarker and therapeutic target.
