Mebendazole inhibits PELI3-mediated ubiquitination of TRADD in non-small cell lung cancer cells

Fannian Li; Xiaoning Li; Haitao Li; Shuai Li; Yanchao Liu; Xianhua Bai; Tianjie Qi; Xiumin Zhao; Yuzheng He

doi:10.3389/fonc.2025.1728170

Mebendazole inhibits PELI3-mediated ubiquitination of TRADD in non-small cell lung cancer cells

Front Oncol. 2026 Jan 9:15:1728170. doi: 10.3389/fonc.2025.1728170. eCollection 2025.

Authors

Fannian Li¹, Xiaoning Li², Haitao Li³, Shuai Li³, Yanchao Liu⁴, Xianhua Bai⁵, Tianjie Qi³, Xiumin Zhao⁶, Yuzheng He⁷

Affiliations

¹ Department of Thoracic Surgery, the First Affiliated Hospital of Xing Tai Medical College, XingTai, Hebei, China.
² Department of Thoracic Surgery, Hebei General Hospital, Shijiazhuang, Hebei, China.
³ The First Department of Pulmonary and Critical Care Medicine, the Second Hospital of Hebei Medical University, Hebei Key Laboratory of Respiratory Critical Care Medicine, Hebei Institute of Respiratory Diseases, Shijiazhuang, Hebei, China.
⁴ The Second Department of Pulmonary and Critical Care Medicine, the Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
⁵ Department of Medical Imaging, the Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
⁶ Department of the Integrated Treatment of Traditional Chinese and Western Medicine, the Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
⁷ Department of Thoracic Surgery, the Second Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.

Abstract

Introduction: Tumor necrosis factor receptor 1-associated death domain protein (TRADD) can trigger proapoptotic autophagy in non-small cell lung cancer (NSCLC). While the potential ubiquitin-protein ligase (E3) against TRADD is not well deciphered.

Methods: UbiBrowser was used to predict the potential E3 ubiquitin ligase to bind with TRADD. Co-immunoprecipitation was performed in HEK293T cells co-transfected with Myc-PELI3, Flag-TRADD, or HA-Ub plasmids. Increasing doses of Myc-PELI3 were transfected into HCC827 and A549 cells, and the relative expression of TRADD was detected. Cycloheximide chase assay was performed in A549 cells transfected with Myc-PELI3 plasmids, and the stability of TRADD was revealed. CCK-8 assay was performed in A549 and HCC827 cells incubated with increasing doses of Mebendazole. The expression of TRADD and PELI3 after Mebendazole incubation was assayed with Western Blot and RT-PCR. The potential E3 ubiquitin ligase of PELI3 was predicted by the UbiBrowser platform, and the binding of PELI3 with TRADD was testified in HEK293T cells co-transfected with Myc-PELI3 and Flag-TRADD plasmids.

Results: PELI3 overexpression diminished the relative protein expression of TRADD, while not affecting the relative mRNA expression in both A549 and HCC827 cells. Cycloheximide assay and following HA-Ub detection demonstrated that PELI3 decreased the protein stability of TRADD by inducing polyubiquitination. Mebendazole inhibited the viability of HCC827 and A549 cells with diminished expression of PELI3 and increased expression of TRADD.

Conclusions: PELI3 can function as an E3 ubiquitin ligase to ubiquitinate TRADD, and Mebendazole might be a promising drug to affect PELI3 expression in NSCLC.

Keywords: E3 ubiquitin ligase; PELI3; TRADD; mebendazole; non-small cell lung cancer.