RNF112-mediated FOXM1 ubiquitination suppresses the proliferation and invasion of gastric cancer
Shengwei Zhang, Jing Wang, Weichao Hu, Lijiao He, Qingyun Tang, Jie Li, Mengmeng Jie, Xinzhe Li, Cheng Liu, Qin Ouyang, Shiming Yang, Changjiang Hu
Shengwei Zhang, Jing Wang, Weichao Hu, Lijiao He, Qingyun Tang, Jie Li, Mengmeng Jie, Xinzhe Li, Cheng Liu, Qin Ouyang, Shiming Yang, Changjiang Hu
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Research Article Gastroenterology Oncology

RNF112-mediated FOXM1 ubiquitination suppresses the proliferation and invasion of gastric cancer

Abstract

Forkhead box M1 (FOXM1) plays a critical role in development physiologically and tumorigenesis pathologically. However, insufficient efforts have been dedicated to exploring the regulation, in particular the degradation of FOXM1. Here, the ON-TARGETplus siRNA library targeting E3 ligases was used to screen potential candidates to repress FOXM1. Of note, mechanism study revealed that RNF112 directly ubiquitinates FOXM1 in gastric cancer, resulting in a decreased FOXM1 transcriptional network and suppressing the proliferation and invasion of gastric cancer. Interestingly, the well-established small-molecule compound RCM-1 significantly enhanced the interaction between RNF112 and FOXM1, which further promoted FOXM1 ubiquitination and subsequently exerted promising anticancer effects in vitro and in vivo. Altogether, we demonstrate that RNF112 suppresses gastric cancer progression by ubiquitinating FOXM1 and highlight the RNF112/FOXM1 axis serves as both prognosis biomarker and therapeutic target in gastric cancer.

Authors

Shengwei Zhang, Jing Wang, Weichao Hu, Lijiao He, Qingyun Tang, Jie Li, Mengmeng Jie, Xinzhe Li, Cheng Liu, Qin Ouyang, Shiming Yang, Changjiang Hu

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Figure 6

RCM-1 enhances the interaction between RNF112 and FOXM1.

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RCM-1 enhances the interaction between RNF112 and FOXM1. (A) Immunoblot ...
(A) Immunoblot analysis of the cellular distribution of FOXM1 and RNF112 in MGC803 cells after DMSO or RCM-1 treatment (10 μM). H3 and tubulin were used as markers of nuclear and cytoplasmic fractions, respectively. (B) Co-IP analysis of the interaction between FOXM1-FLAG and RNF112-HA in HEK293T cells in the presence of DMSO or RCM-1 (10 μM). (C) Ubiquitination of FOXM1 was analyzed after transfection with RNF112 or control in HEK293T cells treated with DMSO or RCM-1 (10 μM). (D) Quantitative reverse transcription–PCR analysis of FOXM1 target genes in MGC803 cells treated with DMSO or RCM-1 (10 μM) (n = 3). (E) Co-IP analysis of RNF112-FOXM1 interaction in HEK293T cells treated with RCM-1 (10 μM) and LMB (25 nM) for 24 hours. (F) Immunoblot analysis of endogenous FOXM1 expression in MGC803 cells treated with RCM-1 (10 μM) and LMB (25 nM) for 24 hours. Ethanol and DMSO were used as vehicle controls. (G) Docking model of the FOXM1/RNF112 complex. Red and purple dashed lines indicate hydrogen bonds and salt bridges, respectively. (H) Bubble plot of binding free energies (ΔGbind) between the key residues of FOXM1 and RNF112. The bubble size corresponds to the absolute value of the binding free energy. (I) Docking model of FOXM1/RCM-1–RNF112/RCM-1 complex (RCM-1 is not shown). (J) Bubble plot of binding free energies between the key residues of FOXM1 and RNF112 in the presence of RCM-1. Blue bubbles indicate the additional key residues. Statistical significance was calculated using Student’s t test (D). *P < 0.05. Complete unedited blots are in the supplemental material. WCL, whole-cell lysate.