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RNF4 silencing induces cell growth arrest and DNA damage by promoting nuclear targeting of p62 in hepatocellular carcinoma

Oncogene volume 41, pages 2275–2286 (2022)Cite this article

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Abstract

Hepatocellular carcinoma (HCC) is one of the largest causes of cancer-related deaths worldwide owing to the limitation of effective treatment options. The ubiquitin-proteasome system has been rapidly recognized as a frequent target of deregulation leading to cancers. Enhanced DNA damage response (DDR) promotes HCC growth and prevents chemosensitivity, and ubiquitin E3 ligases are key modulators in DDR. Therefore, a better understanding of how E3 ligases regulate cell growth and DNA damage may provide novel insights in understanding the oncogenic mechanism and improving the efficacy of DNA damage therapeutic agents. Here, we performed a high-content RNAi screening targeting 52 DDR-related E3 ligases in HCC and found that ring finger protein 4 (RNF4) was essential for HCC growth. RNF4 was highly expressed in HCC tissues, and the expression levels of RNF4 were associated with poor outcomes. RNF4 silencing significantly suppressed the cell growth, and subsequently induced G2/M arrest and apoptosis of HCC cells in vitro; RNF4 silencing also demonstrated the tumor-suppressive efficacy on HCC in vivo. Moreover, RNF4 silencing increased DNA damage, and rendered HCC cells more sensitive to DNA damage drugs and radiation. We found RNF4 functionally interacts with p62, and mechanistic analyses indicated that RNF4 silencing triggered the nuclear enrichment of p62. Moreover, the p62 nuclear targeting was required for increased DNA damage and growth suppression mediated by RNF4 silencing. Thus, our findings suggest RNF4 is essential for HCC proliferation via preventing nuclear translocation of p62. RNF4 silencing promotes DNA damage and may serve as a novel strategy to suppress cell growth and increase the sensitivity of DNA damage therapeutic agents in HCC.

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Fig. 1: Oncogenic role of RNF4 in HCC.
Fig. 2: RNF4 silencing induces G2/M cell cycle arrest and apoptosis.
Fig. 3: RNF4 silencing causes endogenous DNA damage and sensitizes the effect of DNA damage reagents.
Fig. 4: RNF4 interacts with p62.
Fig. 5: RNF4 silencing-induced cell growth arrest and DNA damage are dependent on p62 expression.
Fig. 6: RNF4 silencing induces the nuclear localization of p62 and nuclear p62 involves in RNF4 silencing-induced cell death and DNA damage.
Fig. 7: RNF4 silencing sensitizes the effect of DNA damage reagents in a p62-dependent manner in vivo.

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Acknowledgements

This work was supported by National Natural Science Foundation Grant of China [81770579, 81830080, 81861168038, 81970506, 81800477]. The authors thank Ying Zhao for p62 WT, p62 â–³UBA, p62 â–³PB1, p62 â–³NES and p62 â–³NLS plasmids. The authors thank Dr. Zhaohui Wu and Dr. Mingfang Lv for revising our manuscript. The authors thank Mengxiao Ge for replenishing our experimental data.

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  1. These authors contributed equally: Bin Lv, Yida Pan.

Authors and Affiliations

  1. Department of Digestive Diseases, Huashan Hospital, Fudan University, 12 Middle Wulumuqi Road, Shanghai, China

    Bin Lv, Yida Pan, Jun Zhang, Dongqin Yang & Jie Liu

  2. Institutes of Biomedical Sciences, School of Basic Medical Sciences, Fudan University, 130 Dong’an Road, Shanghai, China

    Daisen Hou, Ping Chen & Jie Liu

  3. Department of Immunology, School of Basic Medical Sciences, Fudan University, 130 Dong’an Road, Shanghai, China

    Daisen Hou, Ping Chen, Yiwei Chu & Jie Liu

  4. Center of Cancer Research, University of Tennessee Health Science Center, Memphis, TN, 38163, USA

    Mingqi Li

  5. College of Veterinary Medicine, Jilin Agricultural University, 2888 Xincheng Street, Changchun, China

    Yan Zeng

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Contributions

BL and YP contributed to acquisition, analysis and interpretation of data, and original draft writing. DH, PC, ML, and ZY contributed to acquisition of data. JZ contributed to analysis and interpretation of data. YC contributed to article revision. DY and JL contributed to study design and article revision.

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Correspondence to Dongqin Yang or Jie Liu.

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Lv, B., Pan, Y., Hou, D. et al. RNF4 silencing induces cell growth arrest and DNA damage by promoting nuclear targeting of p62 in hepatocellular carcinoma. Oncogene 41, 2275–2286 (2022). https://doi.org/10.1038/s41388-022-02247-4

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