Epstein-Barr virus protein boosts cancer gene activity in HPV-positive cervical cells

Scientists have shown that the Epstein-Barr virus nuclear antigen EBNA1 is able to enhance the expression of two cellular genes associated with tumor progression at once - Derlin1 (DERL1) and PSMD10 ( gankyrin ) - in the HeLa model cervical cancer line. In the experiment, three groups of cells were compared: with EBNA1 expression, with an "empty" control plasmid and without transfection. After isolating RNA, the mRNA level was measured by RT-qPCR and the differences were assessed (Mann-Whitney, p < 0.05). Result: against the background of EBNA1, DERL1 increased approximately 3-fold (p ≈ 0.028), PSMD10 - approximately 2-fold (p ≈ 0.02-0.03); changes in ZEB1 and CNN3 were statistically insignificant. The authors suggested that such transcriptional 'twisting' may support tumor cell survival and accelerate cervical cancer progression, especially in the context of possible HPV+EBV co-infection.

Background of the study

Cervical cancer is almost always associated with the persistence of high-oncogenic HPV types, but increasing evidence suggests the role of co-infections in accelerating malignancy and progression. Epstein-Barr virus (EBV) is frequently found in cervical tissues in association with HPV; meta-reviews and case series have linked this “duet” to higher grades of dysplasia and adverse molecular features. It is suggested that EBV may enhance HPV transformation signals, increase cellular resistance to apoptosis, and remodel the tumor microenvironment.

The key EBV latency protein, EBNA1, is present in nearly all EBV-associated tumors. It maintains episome copy number, regulates viral promoters, and is capable of influencing cellular gene transcription, thereby fine-tuning host expression networks. These properties make EBNA1 a likely cofactor in carcinogenesis and a candidate target for interventions in EBV-positive epithelial tumors.

Genes associated with protein quality control and degradation are considered as potential "nodes" of such an effect. Derlin1 (DERL1) is a component of the ERAD (ER removal of misfolded proteins) pathway, which, when hyperactivated, supports the survival of stressed cancer cells; PSMD10 (gankyrin) is a regulatory subunit of the proteasome, known to suppress the p53/RB pathway and enhance proliferation. Both genes have been repeatedly described as pro-oncogenic in various models and are therefore of interest as readers of the effect of EBV on the cellular environment.

Against this background, a new study in Genes & Cancer tests whether EBNA1 can directly “tune” the expression of DERL1 and PSMD10 in cervical cancer cells (HeLa model) and thereby contribute to the survival/resistance phenotype. The authors evaluate the mRNA levels of the target genes compared to controls and discuss the resulting signaling pattern as a possible molecular signature of EBV during co-infection with HPV.

Why is this important?

HPV is the main etiologic factor of cervical cancer, but additional “cofactors” are often required for malignant transformation. EBV is known as an oncovirus in epithelial and lymphoid tumors and is often found in cervical tissues together with HPV. By showing that one of the key latent proteins of EBV directly shifts the expression of genes involved in ER stress (DERL1) and regulation of the proteasome/p53-RB pathway (gankyrin, PSMD10), our work raises a practical question: can EBV enhance the resistance of cervical cells to apoptosis and treatment, making the tumor more “viable”?

Who exactly does EBNA1 “touch”?

• DERL1 (Derlin1) is a membrane protein of the endoplasmic reticulum, a participant in the removal of misfolded proteins (ERAD) system; its overexpression is associated with cell growth/migration and resistance to apoptosis.
• PSMD10 (gankyrin) is a regulatory subunit of the 26S proteasome; it negatively affects p53 and RB1 via the MDM2/CDK4 axis, supporting tumor proliferation and survival.
• ZEB1 is a transcriptional regulator of epithelial-mesenchymal transition (EMT); increased expression often accompanies invasion and drug resistance.
• CNN3 - actin-related calponin-3; associated with migration/metastasis in some solid tumors.

How to read this data without unnecessary hype

This is an in vitro model on one cell line (HeLa), measurements are only at the mRNA level without confirmation by protein tests (Western, immunohistochemistry), functional consequences (proliferation, invasion, apoptosis) were not tested. There is no comparison with normal cervical epithelium and with other HPV statuses, which means that the transferability of the results to the clinic is limited. This makes a clear "signal" for two genes even more valuable: it outlines the molecular trace of EBNA1, which is worth checking in co-infection models of HPV/EBV, on primary material and in animal experiments.

What this might mean next - practical implications and hypotheses

• Co-infection biomarkers: The combination of HPV profile with DERL1/PSMD10 expression and EBV markers may help to identify risk subgroups and predict resistance to therapy.
• Therapeutic entry points: If EBNA1 consistently upregulates DERL1/PSMD10, it is logical to test inhibitors of the ER stress/ERAD pathway and modifiers of the proteasome/p53-MDM2 axis in an EBV-positive context.
• Diagnostic stratification. In clinical series of cervical cancer, it is worthwhile to correlate EBV status with treatment outcomes and DERL1/PSMD10 expression to understand who has an additional “viral contribution” to aggressiveness.

The limitations are acknowledged by the authors - and that's a plus

• One HeLa line; no HPV positive/negative line panels.
• No protein and functional confirmation (RT-qPCR only).
• The study is associational: the mechanism of direct regulation of EBNA1 promoters for these genes has not been shown, as well as the dependence of the effects on dose/time.
• Validation on clinical cervical samples has not been performed.

Conclusion

The work neatly adds a new building block to our understanding of possible HPV-EBV cooperation in cervical cancer: the latent protein EBNA1 is able to shift the expression of genes that support tumor cell survival and proliferation. Clinical implications are still a long way off, but DERL1 and PSMD10 look like reasonable candidates for indicators and targets in the EBV-positive context – provided that the results are confirmed at the protein level, function, and in real tumor tissues.

Source: Alipour AH, Hashemi SMA, Gharahkhani F., Katanchi A., Farhadi A., Sarvari J. Epstein-Barr virus nuclear antigen 1 upregulates Derlin1 and PSMD10 expression in HeLa cells. Genes & Cancer (accepted 24 July 2025; published 6 August 2025). https://doi.org/10.18632/genesandcancer.242