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Insufficient ablation induces E3-ligase Nedd4 to promote hepatocellular carcinoma progression by tuning TGF-β signaling

Oncogene volume 41, pages 3197–3209 (2022)Cite this article

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Abstract

Thermal ablation is a main curative therapy for early-stage hepatocellular carcinoma (HCC). However, insufficient ablation has been shown to promote HCC progression. E3 ligases have been approved to play important roles in malignant tumors. Whether E3 ligases are involved in HCC progression caused by insufficient ablation remains unclear. Herein, using RNA-sequencing coupled with an in vitro loss-of-function screen, we found that the E3 ligase Neuronal Precursor cell-expressed Developmentally Downregulated 4 (Nedd4) was upregulated in HCC insufficient ablation tissues and promoted HCC cells migration. The upregulation of Nedd4 was induced by METTL14-mediated N6-methyladenosine modification after sublethal heat treatment. Knockdown of Nedd4 inhibited HCC metastasis and growth in vitro and in vivo. Mechanistically, Nedd4 enhanced TGF-β signal transduction mediated tumor progression by directly binding to TGF-β type I receptor (TGFBR1) and forming K27-linked ubiquitin at Lysine 391. Additionally, the adverse effect on HCC of sublethal heat treatment was mediated by Nedd4. Clinically, high Nedd4 expression was positively correlated with aggressive tumor phenotypes and poor prognosis in HCC patients. Patient-derived xenograft (PDX) model confirmed this conclusion. Collectively, this study demonstrated that Nedd4 induced by insufficient ablation plays a crucial role in promoting HCC progression and provides a novel therapeutic target for HCC.

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Fig. 1: Nedd4 was induced by insufficient ablation in HCC.
Fig. 2: Knockdown of Nedd4 inhibited HCC metastasis and growth.
Fig. 3: Overexpression of wild type Nedd4 rescued HCC progression in Nedd4 knockdown cells.
Fig. 4: Nedd4 enhanced EMT through TGF-β/smad2/3 signaling.
Fig. 5: Nedd4 directly recognized TGFBR1 through its WW domain and ubiquitinated K27-poly-ubiquitin chain on TGFBR1 Lysine 391.
Fig. 6: HCC cell metastasis is promoted by sublethal heat stress in a Nedd4-dependent way.
Fig. 7: Clinical significance of Nedd4 inhibition in HCC.
Fig. 8: Working model.

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All data are available in the main text or the supplementary materials.

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Funding

National Natural Science Foundation of China (No. 81772625, No. 81972301 and No. 81802421). Guangzhou Science and Technology Program of China (No. 201804020093).

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Author notes

  1. These authors contributed equally: Kai Li, Yi Niu, Yichuan Yuan, Jiliang Qiu.

Authors and Affiliations

  1. State Key Laboratory of Oncology in South China and Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center, Guangzhou, PR China

    Kai Li, Yi Niu, Yichuan Yuan, Jiliang Qiu, Yunxing Shi, Chengrui Zhong, Zhiyu Qiu, Keren Li, Zhu Lin, Zhenkun Huang, Chao Zhang, Dinglan Zuo, Wei He, Yunfei Yuan & Binkui Li

  2. Department of Liver Surgery, Sun Yat-sen University Cancer Center, Guangzhou, PR China

    Kai Li, Yichuan Yuan, Jiliang Qiu, Yunxing Shi, Chengrui Zhong, Zhiyu Qiu, Keren Li, Zhu Lin, Zhenkun Huang, Wei He, Yunfei Yuan & Binkui Li

  3. Department of Pathology, Sun Yat-sen University Cancer Center, Guangzhou, PR China

    Chao Zhang

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Contributions

Conceptualization: BL and YY. Methodology: KL, YN, YY and YS, Investigation: CZ and ZQ, Visualization: ZH and DZ. Statistical analysis: ZL and KL. Supervision: BL, YY, and CZ. Writing—original draft: KL and YN. Writing—review & editing: BL, WH, and JQ.

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Correspondence to Yunfei Yuan or Binkui Li.

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This study complied with the standards of the 1975 Declaration of Helsinki and the experiments were approved by the Ethics Committee of Sun Yat-sen University Cancer Center (Approval No: GZR2017-116). The animal experiments were approved by the institutional ethics committee of Sun Yat-sen University, Guangzhou, China (Approval No: L102012019120E).

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Li, K., Niu, Y., Yuan, Y. et al. Insufficient ablation induces E3-ligase Nedd4 to promote hepatocellular carcinoma progression by tuning TGF-β signaling. Oncogene 41, 3197–3209 (2022). https://doi.org/10.1038/s41388-022-02334-6

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