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Romidepsin hepatocellular carcinoma suppression in mice is associated with deregulated gene expression of bone morphogenetic protein and Notch signaling pathway components

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Abstract

Recently, our group showed that Romidepsin, a histone deacetylase inhibitor (HDACi), suppressed diethylnitrosamine (DEN)-induced hepatocellular carcinoma (HCC) in mice. In the present study, we investigated the effect of Romidepsin-treatment on gene expression levels of components of Bmp and Notch signaling pathways, which are both known to be aberrantly regulated in hepatocarcinogenesis. Total RNA from liver tissue samples and paraffin-embedded livers were retrieved from a recent experiment where C57BL/6 mice were treated with Romidepsin 10 months after DEN challenge and sacrificed 2 months later. RT qPCR was used for quantification of gene expression and immunohistochemistry for in situ protein detection. Regarding Bmp pathway, Romidepsin HCC-suppression was found to correlate significantly with Bmp2 and Bmp7 ligand up- and down-regulation, respectively. Intracellularly, Romidepsin-treated HCC mice exhibited a significant elevation of Bmp-inhibitor Smurf2 and Bmp-target gene Id3, as compared to the HCC untreated controls. Concerning Notch signaling, higher expression levels of ligands Jag1/Dll4, accompanied by a decreased expression of receptor Notch2, were identified in the Romidepsin-treated group. Τhe anti-oncogenic effect of Romidepsin, also correlated significantly with an increased expression of Hes1 target, as well as an up- and down-regulation of Klf4 and Sox9 transcription factors, respectively. Moreover, the cancer-related genes Snai2 and p21, known to be involved in many signaling pathways, including Bmp and Notch, were also found to be downregulated in Romidepsin-treated mice. Romidepsin HCC suppression is associated with gene expression deregulation of selective components of both Bmp and Notch signaling cascades.

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Abbreviations

Bambi:

Bmp and activin membrane-bound inhibitor

Bim:

BCL2-like 11

Bmp:

Bone morphogenetic proteins

Chrd:

Chordin

Co-Smad:

Common-mediator Smad

DEN:

Diethylnitrosamine

Dll:

Delta ligand

Grem1:

Gremlin 1

HAT:

Histone acetyltransferases

HDAC:

Histone deacetylases

HDACi:

Histone deacetylase inhibitors

Hes1:

Hairy/enhancer of split 1

Hey1:

Hes-related 1

Id:

Inhibitor of DNA binding

I-Smads:

Inhibitory Smads

Jag:

Jagged

Klf4:

Kruppel-like factor 4

NICD:

Notch intracellular domain

Nog:

Noggin

p300:

E1A binding protein p300

R-Smads:

Receptor-regulated Smads

Runx:

Runt-domain transcription factors

Ski:

SKI family transcriptional corepressor

Smurfs:

Smad-ubiquitination regulatory factors

Snai2:

Snail family zinc finger 2

SnoN:

SKI-like

Sox9:

Sex determining region Y-box 9

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Acknowledgements

The authors are grateful to the Bodossaki Foundation for the kind donation of the real-time PCR instrumentation.

Funding

This work was funded as Scholarship by the Experimental, Educational and Research Center ELPEN Pharmaceuticals.

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Authors and Affiliations

  1. Laboratory of Biochemistry and Toxicology, School of Veterinary Medicine, Faculty of Health Sciences, Aristotle University of Thessaloniki, 54124, Thessaloniki, Greece

    Hara Afaloniati & Katerina Angelopoulou

  2. Laboratory of Pathology, School of Veterinary Medicine, Faculty of Health Sciences, Aristotle University of Thessaloniki, Thessaloniki, Greece

    Theofilos Poutahidis

  3. First Department of Surgery, Medical School, General Hospital Papageorgiou, Aristotle University of Thessaloniki, Thessaloniki, Greece

    Alexander Giakoustidis, Athanasios Gargavanis & Dimitrios Giakoustidis

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  1. Hara Afaloniati
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Correspondence to Katerina Angelopoulou.

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Afaloniati, H., Poutahidis, T., Giakoustidis, A. et al. Romidepsin hepatocellular carcinoma suppression in mice is associated with deregulated gene expression of bone morphogenetic protein and Notch signaling pathway components. Mol Biol Rep 48, 551–562 (2021). https://doi.org/10.1007/s11033-020-06089-9

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