Objective: Transcribed-ultraconserved regions (T-UCR) are long non-coding RNAs which are conserved across species and are involved in carcinogenesis. We studied T-UCRs downstream of the Wnt/β-catenin pathway in liver cancer. Design: Hypomorphic Apc mice (Apcfl/fl) and thiocetamide (TAA)-treated rats developed Wnt/β-catenin dependent hepatocarcinoma (HCC) and cholangiocarcinoma (CCA), respectively. T-UCR expression was assessed by microarray, real-time PCR and in situ hybridisation. Results: Overexpression of the T-UCR uc.158-could differentiate Wnt/β-catenin dependent HCC from normal liver and from β-catenin negative diethylnitrosamine (DEN)-induced HCC. uc.158-was overexpressed in human HepG2 versus Huh7 cells in line with activation of the Wnt pathway. In vitro modulation of β-catenin altered uc.158-expression in human malignant hepatocytes. uc.158-expression was increased in CTNNB1-mutated human HCCs compared with non-mutated human HCCs, and in human HCC with nuclear localisation of β-catenin. uc.158-was increased in TAA rat CCA and reduced after treatment with Wnt/β-catenin inhibitors. uc.158-expression was negative in human normal liver and biliary epithelia, while it was increased in human CCA in two different cohorts. Locked nucleic acid-mediated inhibition of uc.158-reduced anchorage cell growth, 3D-spheroid formation and spheroid-based cell migration, and increased apoptosis in HepG2 and SW1 cells. miR-193b was predicted to have binding sites within the uc.158-sequence. Modulation of uc.158-changed miR-193b expression in human malignant hepatocytes. Co-transfection of uc.158-inhibitor and anti-miR-193b rescued the effect of uc.158-inhibition on cell viability. Conclusions: We showed that uc.158-is activated by the Wnt pathway in liver cancers and drives their growth. Thus, it may represent a promising target for the development of novel therapeutics.

Wnt signalling modulates transcribed-ultraconserved regions in hepatobiliary cancers

NEGRINI, Massimo;
2017

Abstract

Objective: Transcribed-ultraconserved regions (T-UCR) are long non-coding RNAs which are conserved across species and are involved in carcinogenesis. We studied T-UCRs downstream of the Wnt/β-catenin pathway in liver cancer. Design: Hypomorphic Apc mice (Apcfl/fl) and thiocetamide (TAA)-treated rats developed Wnt/β-catenin dependent hepatocarcinoma (HCC) and cholangiocarcinoma (CCA), respectively. T-UCR expression was assessed by microarray, real-time PCR and in situ hybridisation. Results: Overexpression of the T-UCR uc.158-could differentiate Wnt/β-catenin dependent HCC from normal liver and from β-catenin negative diethylnitrosamine (DEN)-induced HCC. uc.158-was overexpressed in human HepG2 versus Huh7 cells in line with activation of the Wnt pathway. In vitro modulation of β-catenin altered uc.158-expression in human malignant hepatocytes. uc.158-expression was increased in CTNNB1-mutated human HCCs compared with non-mutated human HCCs, and in human HCC with nuclear localisation of β-catenin. uc.158-was increased in TAA rat CCA and reduced after treatment with Wnt/β-catenin inhibitors. uc.158-expression was negative in human normal liver and biliary epithelia, while it was increased in human CCA in two different cohorts. Locked nucleic acid-mediated inhibition of uc.158-reduced anchorage cell growth, 3D-spheroid formation and spheroid-based cell migration, and increased apoptosis in HepG2 and SW1 cells. miR-193b was predicted to have binding sites within the uc.158-sequence. Modulation of uc.158-changed miR-193b expression in human malignant hepatocytes. Co-transfection of uc.158-inhibitor and anti-miR-193b rescued the effect of uc.158-inhibition on cell viability. Conclusions: We showed that uc.158-is activated by the Wnt pathway in liver cancers and drives their growth. Thus, it may represent a promising target for the development of novel therapeutics.
GUT
Carotenuto, Pietro; Fassan, Matteo; Pandolfo, Rosantony; Lampis, Andrea; Vicentini, Caterina; Cascione, Luciano; Paulus Hock, Viola; Boulter, Luke; Guest, Rachel; Quagliata, Luca; Hahne, Jens Claus; Ridgway, Rachel; Jamieson, Tam; Athineos, Dimitris; Veronese, Angelo; Visone, Rosa; Murgia, Claudio; Ferrari, Giulia; Guzzardo, Vincenza; Evans, Thomas Ronald Jeffry; Macleod, Martin; Feng, Gui Ji; Dale, Trevor; Negrini, Massimo; Forbes, Stuart J.; Terracciano, Luigi; Scarpa, Aldo; Patel, Tushar; Valeri, Nicola; Workman, Paul; Sansom, Owen; Braconi, Chiara
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11392/2368232
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