Decitabine suppresses tumor growth by activating mouse mammary tumor virus and interferon-β pathways

Ryan Johnson; Andrew Brola; Cade Wycoff; William Wycoff; Seth Neumeyer; Richard Tuttle; Sarah Light; Jiayi Li; Stephen Christensen; Yingguang Liu

doi:10.17305/bb.2025.12852

Authors

Ryan Johnson Department of Biomedical Sciences, College of Osteopathic Medicine, Liberty University, Lynchburg, VA, USA
Andrew Brola Department of Biomedical Sciences, College of Osteopathic Medicine, Liberty University, Lynchburg, VA, USA https://orcid.org/0009-0005-0033-5526
Cade Wycoff Department of Biomedical Sciences, College of Osteopathic Medicine, Liberty University, Lynchburg, VA, USA
William Wycoff Department of Biomedical Sciences, College of Osteopathic Medicine, Liberty University, Lynchburg, VA, USA
Seth Neumeyer Department of Biomedical Sciences, College of Osteopathic Medicine, Liberty University, Lynchburg, VA, USA https://orcid.org/0009-0003-2251-9444
Richard Tuttle Department of Biomedical Sciences, College of Osteopathic Medicine, Liberty University, Lynchburg, VA, USA
Sarah Light Department of Biomedical Sciences, College of Osteopathic Medicine, Liberty University, Lynchburg, VA, USA
Jiayi Li Department of Biomedical Sciences, College of Osteopathic Medicine, Liberty University, Lynchburg, VA, USA
Stephen Christensen Department of Biomedical Sciences, College of Osteopathic Medicine, Liberty University, Lynchburg, VA, USA
Yingguang Liu Department of Biomedical Sciences, College of Osteopathic Medicine, Liberty University, Lynchburg, VA, USA

DOI:

https://doi.org/10.17305/bb.2025.12852

Keywords:

Decitabine , DNA methyltransferase inhibitor, mouse mammary tumor virus, interferon, tumor, cancer, 4T1, MC38, interferon regulatory factor 7

Abstract

Decitabine (DAC), a DNA methyltransferase inhibitor (DNMTi), is clinically effective in hematological malignancies such as myelodysplastic syndrome and acute myeloid leukemia, but its precise antineoplastic mechanisms remain incompletely understood. Beyond promoter demethylation, DAC is known to activate endogenous retroviruses (ERVs) and trigger type I interferon (IFN-I) responses, a phenomenon known as viral mimicry. The aim of this study was to investigate the roles of the mouse mammary tumor virus (MMTV) and interferon-β (IFN-β) in DAC-mediated tumor suppression. We employed two murine tumor models—4T1 mammary carcinoma and MC38 colon adenocarcinoma—in syngeneic immunocompetent mice, immunodeficient nude mice, and in vitro cultures. RNA and protein expression were assessed by quantitative PCR and immunoblotting, while functional contributions of MMTV and IFN-β were tested using short hairpin RNA (shRNA) knockdowns. DAC treatment suppressed tumor growth and pulmonary metastasis in vivo and inhibited cancer cell proliferation in vitro. It induced transcription of MMTV and expression of IFN-β, with a strong negative correlation between MMTV Env protein levels and tumor mass. Knockdown of either MMTV or IFN-β conferred resistance to DAC, confirming their functional roles. Reciprocal regulation was observed: MMTV knockdown reduced IFN-β expression, while IFN-β knockdown increased MMTV Env accumulation. Furthermore, DAC upregulated interferon regulatory factor 7 (IRF7), but this effect declined during prolonged treatment, suggesting a temporally restricted therapeutic window. In conclusion, our findings provide in vivo support for the viral mimicry hypothesis and demonstrate that MMTV and IFN-β contribute to DAC-mediated tumor suppression. The observed IRF7 downregulation and potential induction of immune checkpoints highlight the importance of therapeutic strategies combining DNMTis with immune checkpoint blockade to sustain antineoplastic efficacy.