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Silencing of RND3/RHOE inhibits the growth of human hepatocellular carcinoma and is associated with reversible senescence

Cancer Gene Therapy volume 29, pages 437–444 (2022)Cite this article

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Abstract

Rnd3/RhoE is an atypical Rho GTPase family member, known to be deregulated in many types of cancer. Previously, we showed that RND3 expression is downregulated in hepatocellular carcinoma (HCC) cell lines and tissues. In cancer cells, Rnd3 is involved in the regulation of cell proliferation and cell invasion. The implication of Rnd3 in HCC invasion was importantly studied whereas its role in cell growth needs further investigation. Thus, in this work, we aimed to better understand the impact of Rnd3 on tumor hepatocyte proliferation. Our results indicate that the silencing of RND3 induces a cell growth arrest both in vitro in 2D and 3D culture conditions and in vivo in tumor xenografts. The growth alteration after RND3 silencing in HCC cells is not due to an increase of cell death but to the induction of senescence. This RND3 knockdown-mediated phenomenon is dependent on the decrease of hTERT expression. Interestingly, after re-expression of RND3, these cells are able to bypass senescence and regain the ability to proliferate, with a re-expression of hTERT. Given that a low expression of Rnd3 is linked to the presence of satellite nodules in HCC, the transient senescence state observed might play a role in cancer progression.

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Fig. 1: RND3 depletion alters Hep3B cell growth in vitro in 2D and 3D and in vivo.
Fig. 2: Silencing of Rnd3 induces an alteration of cell proliferation in HCC cells.
Fig. 3: Rnd3 protein knockdown induces senescence in Hep3B cells associated with the downregulation of hTERT.
Fig. 4: The overexpression of hTERT after Rnd3 KD reverses the senescence phenotype in Hep3B cell line and the re-expression of RND3 allows cells to escape from senescence state.

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Acknowledgements

We thank V. Guyonnet-Duperat and V. Pitard from the vectorology and flow cytometry core facilities, respectively (TBMCore, UMS005, Bordeaux). We thank C. Péanne for her help with apoptosis analyses. SB and SS were supported by postdoctoral fellowships from, respectively, the Fondation ARC/Région Nouvelle-Aquitaine and the Fondation pour la Recherche Médicale. L. Paysan was supported by a PhD fellowship from the Région Aquitaine. This work was supported by grants from La Ligue contre le Cancer (comité régional) and from Institut National du Cancer (PLBIO-INCa2014-182) (to VM). VM is supported by La Fondation pour la Recherche Médicale “Equipe labellisée 2018”.

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Authors and Affiliations

  1. Univ. Bordeaux, INSERM, BaRITOn, U1053, F-33000, Bordeaux, France

    Sara Basbous, Lisa Paysan, Sandra Sena, Nathalie Allain, Edith Chevret, Valérie Lagrée & Violaine Moreau

  2. Hepatology Unit, Bordeaux University Hospital, Bordeaux, France

    Jean-Baptiste Hiriart

  3. Histopathology Platform, TBM-Core US 005, F-33076, Bordeaux, France

    Nathalie Dugot-Senant

  4. Animal Core Facility, Univ. Bordeaux, F-33000, Bordeaux, France

    Benoît Rousseau

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  1. Sara Basbous
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  9. Valérie Lagrée
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  10. Violaine Moreau
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Contributions

Study design: SB, EC, VL, VM. Generation of experimental data: SB, LP, SS, NA, J-BH, ND-S, BR. Analysis and interpretation of data: SB, SS, LP, NA, J-BH, VM. Writing of the manuscript: SB, VM. Critical reading of the manuscript: EC, VL. Supervision of the project: VM.

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Correspondence to Violaine Moreau.

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Basbous, S., Paysan, L., Sena, S. et al. Silencing of RND3/RHOE inhibits the growth of human hepatocellular carcinoma and is associated with reversible senescence. Cancer Gene Ther 29, 437–444 (2022). https://doi.org/10.1038/s41417-022-00445-6

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