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Open AccessDOI: 10.3724/abbs.2025048Original Research

Withaferin A combined with ricolinostat: a potent synergistic therapy for cervical cancer through regulating p53 ubiquitination and acetylation

🇨🇳 Original Chinese Title: Withaferin A combined with ricolinostat: a potent synergistic therapy for cervical cancer through regulating p53 ubiquitination and acetylation

Tian Chen¹,Yiting Xu¹,Kunming Yang¹,Yutong Du¹,Zhuan Zhu¹,Lingling Xu¹,Xinrong Wang¹,Yi Yin¹,Yu Hu¹,Chengcheng Wang¹,Ronggui Hu¹,Chuanyin Li¹

Medical College of Guizhou University, Guiyang 550025, China

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Withaferin A combined with ricolinostat: a potent synergistic therapy for cervical cancer through regulating p53 ubiquitination and acetylation
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Acta Biochimica et Biophysica Sinica
Published:2025Edition:Vol. 57, Issue 12 • pp. 2133-2136Citation:Tian Chen et al. (2025), Acta Biochimica et Biophysica Sinica
Impact FactorPremier Chinese Biomedical Journal indexed in SinoBioData: Acta Biochimica et Biophysica Sinica (生物化学与生物物理学报).
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Key Takeaways & Executive Findings

  • • The combination of Withaferin A (WA) and ricolinostat (RIC) synergistically inhibits cervical cancer cell proliferation and colony formation in vitro, with a combination index (CI) < 1. • In vivo, the WA+RIC combination significantly reduces tumor volume and weight in a HeLa xenograft mouse model compared to single-agent treatments. • The synergistic effect is attributed to enhanced p53 stability via inhibition of ubiquitination (by WA) and acetylation (by RIC), restoring p53 tumor-suppressive function. • This study provides a promising therapeutic strategy for cervical cancer, particularly for HPV-positive cases where p53 is degraded by E6AP.
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Abstract

As a classic tumor suppressor gene, p53 has been extensively studied since its discovery in the mid-1980s. Research findings have revealed that p53 protein expression is suppressed in various cancers [1]. For example, in cervical cancer, p53 predominantly exists in a wild-type form to maintain its biological function [2]. Nevertheless, its tumor-suppressive activity is significantly impaired because of rapid protein degradation, short half-life, and low levels. Post-translational modifications (PTMs) of p53, such as ubiquitination, acetylation, phosphorylation and methylation, are critical regulators of its stability, activity, conformation, localization, and interactions with cofactors [3]. Among these, ubiquitination and acetylation play central roles in controlling p53 protein stability and activity [4]. Therefore, targeting p53 PTMs to modulate its ubiquitination and acetylation levels represents an effective strategy to increase its stability and tumor-suppressive function, offering a promising avenue for cervical cancer drug development. In 99% of cervical cancers (high-risk human papillomavirus-positive), E3 ubiquitin ligase E6-associated protein (E6AP) mediates the ubiquitination degradation of p53 [5], whereas histone deacetylase 6 (HDAC6) deacetylates p53. In this study, we explored the possibility of combining the natural product withferin A (WA) with the HDAC6 inhibitor ricolinostat (RIC) to treat cervical cancer cells, with a focus on the ubiquitination and acetylation of p53 and the consequences for its stability. These results suggested that the combination of WA and RIC is more effective than either treatment alone in inhibiting the degradation and increasing the stability of p53, thereby synergistically slowing the onset and progression of cervical cancer.

1. Introduction

As a classic tumor suppressor gene, p53 has been extensively studied since its discovery in the mid-1980s. Research findings have revealed that p53 protein expression is suppressed in various cancers [1]. For example, in cervical cancer, p53 predominantly exists in a wild-type form to maintain its biological function [2]. Nevertheless, its tumor-suppressive activity is significantly impaired because of rapid protein degradation, short half-life, and low levels.

Post-translational modifications (PTMs) of p53, such as ubiquitination, acetylation, phosphorylation and methylation, are critical regulators of its stability, activity, conformation, localization, and interactions with cofactors [3]. Among these, ubiquitination and acetylation play central roles in controlling p53 protein stability and activity [4]. Therefore, targeting p53 PTMs to modulate its ubiquitination and acetylation levels represents an effective strategy to increase its stability and tumor-suppressive function, offering a promising avenue for cervical cancer drug development.

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Cite This Research Paper
Tian Chen, Yiting Xu, Kunming Yang, Yutong Du, Zhuan Zhu, Lingling Xu, Xinrong Wang, Yi Yin, Yu Hu, Chengcheng Wang, Ronggui Hu, Chuanyin Li (2026). Withaferin A combined with ricolinostat: a potent synergistic therapy for cervical cancer through regulating p53 ubiquitination and acetylation. Acta Biochimica et Biophysica Sinica. https://doi.org/10.3724/abbs.2025048
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Frequently Asked Questions

What is the mechanism of action of the combination therapy?

The combination of Withaferin A (WA) and ricolinostat (RIC) synergistically enhances p53 stability by inhibiting its ubiquitination and acetylation, respectively. WA likely inhibits E6AP-mediated ubiquitination, while RIC inhibits HDAC6-mediated deacetylation, leading to increased p53 levels and tumor-suppressive activity.

How effective is the combination therapy in preclinical models?

In vitro, the combination significantly reduces cell viability and colony formation in cervical cancer cell lines (HeLa, SiHa, Caski) with a combination index < 1, indicating synergy. In vivo, it significantly reduces tumor volume and weight in a HeLa xenograft mouse model compared to single agents.

What are the potential clinical implications of this study?

This study provides a promising therapeutic strategy for cervical cancer, particularly for HPV-positive cases where p53 is degraded by E6AP. The combination of WA and RIC could be developed as a novel treatment option to restore p53 function and improve patient outcomes.

Are there any known side effects of the combination therapy?

The study did not report specific side effects, but as a preclinical study, further toxicity and safety evaluations are needed. The doses used in mice (3 mg/kg WA and 10 mg/kg RIC) were well-tolerated, but clinical trials are required to assess safety in humans.

What are the next steps for this research?

Future research should focus on elucidating the detailed molecular mechanisms, optimizing dosing regimens, and conducting pharmacokinetic and toxicological studies. Ultimately, clinical trials are needed to validate the efficacy and safety of the combination therapy in cervical cancer patients.

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