Identification of DUSP4/6 overexpression as a potential rheostat to NRAS-induced hepatocarcinogenesis

URN zum Zitieren dieses Dokuments:: urn:nbn:de:bvb:355-epub-550219
DOI zum Zitieren dieses Dokuments:: 10.5283/epub.55021

Klemm, Sophie ; Evert, Katja

; Utpatel, Kirsten

; Muggli, Alexandra ; Simile, Maria M. ; Chen, Xin ; Evert, Matthias ; Calvisi, Diego F.

; Scheiter, Alexander

Lizenz: Creative Commons Namensnennung 4.0 International
PDF - Veröffentlichte Version
(4MB)

Veröffentlichungsdatum dieses Volltextes: 17 Nov 2023 08:57

Alternative Links zum Volltext:DOI Verlag

Details

Vorschau

Bibliographische Daten exportieren

Zusammenfassung

Zusammenfassung

Background
Upregulation of the mitogen-activated protein kinase (MAPK) cascade is common in hepatocellular carcinoma (HCC). Neuroblastoma RAS viral oncogene homolog (NRAS) is mutated in a small percentage of HCC and is hitherto considered insufficient for hepatocarcinogenesis. We aimed to characterize the process of N-Ras-dependent carcinogenesis in the liver and to identify potential therapeutic vulnerabilities.
Methods
NRAS V12 plasmid was delivered into the mouse liver via hydrodynamic tail vein injection (HTVI). The resulting tumours, preneoplastic lesions, and normal tissue were characterized by NanoString® gene expression analysis, Western Blot, and Immunohistochemistry (IHC). The results were further confirmed by in vitro analyses of HCC cell lines.
Results
HTVI with NRAS V12 plasmid resulted in the gradual formation of preneoplastic and neoplastic lesions in the liver three months post-injection. These lesions mostly showed characteristics of HCC, with some exceptions of spindle cell/ cholangiocellular differentiation. Progressive upregulation of the RAS/RAF/MEK/ERK signalling was detectable in the lesions by Western Blot and IHC. NanoString® gene expression analysis of preneoplastic and tumorous tissue revealed a gradual overexpression of the cancer stem cell marker CD133 and Dual Specificity Phosphatases 4 and 6 (DUSP4/6). In vitro, transfection of HCC cell lines with NRAS V12 plasmid resulted in a coherent upregulation of DUSP4 and DUSP6. Paradoxically, this upregulation in PLC/PRF/5 cells was accompanied by a downregulation of phosphorylated extracellular-signal-regulated kinase (pERK), suggesting an overshooting compensation. Silencing of DUSP4 and DUSP6 increased proliferation in HCC cell lines.
Conclusions
Contrary to prior assumptions, the G12V NRAS mutant form is sufficient to elicit hepatocarcinogenesis in the mouse. Furthermore, the upregulation of the MAPK cascade was paralleled by the overexpression of DUSP4, DUSP6, and CD133 in vivo and in vitro. Therefore, DUSP4 and DUSP6 might fine-tune the excessive MAPK activation, a mechanism that can potentially be harnessed therapeutically.

Nur für Besitzer und Autoren: Kontrollseite des Eintrags

Downloadstatistik

Altmetric

Alternative Statistik (altmetrics)

Weitere Literatur (mittels CORE)

Details

Vorschau

Bibliographische Daten exportieren

Zusammenfassung

Zusammenfassung

Downloadstatistik

Downloads

Alternative Statistik (altmetrics)

Weitere Literatur (mittels CORE)

Universitätsbibliothek