Hepatitis C virus proteins modulate microRNA expression and chemosensitivity in malignant hepatocytes

Chiara Braconi, Nicola Valeri, Pierluigi Gasparini, Nianyuan Huang, Cristian Taccioli, Gerard Nuovo, Tetsuro Suzuki, Carlo Maria Croce, Tushar C Patel

Research output: Contribution to journalArticle

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Abstract

Purpose: Hepatocellular cancer (HCC) is highly resistant to chemotherapy and is associated with poor prognosis. Chronic hepatitis C virus (HCV) infection is a major cause of HCC. However, the effect of viral proteins in mediating chemosensitivity in tumor cells is unknown. We postulated that HCV viral proteins could modulate therapeutic responses by altering host cell microRNA (miRNA) expression. Experimental Design: HepG2 malignant hepatocytes were stably transfected with full-length HCV genome (Hep-394) or an empty vector (Hep-SWX). MiRNA profiling was done by using a custom microarray, and the expression of selected miRNAs was validated by real-time PCR. Protein expression was assessed by Western blotting, whereas caspase activation was assessed by a luminometric assay. Results: The IC50 to sorafenib was lower in Hep-394 compared with Hep-SWX control cells. Alterations in miRNA expression occurred with 10 miRNAs downregulated >2-fold and 23 miRNAs upregulated >2-fold in Hep-394 cells compared with controls. Of these, miR-193b was overexpressed by 5-fold in Hep-394 cells. miR-193b was predicted to target Mcl-1, an antiapoptotic protein that can modulate the response to sorafenib. The expression of Mcl-1 was decreased, and basal caspase-3/7 activity and poly ADP ribose polymerase cleavage were increased in Hep-394 cells compared with controls. Moreover, transfection with precursors to miR-193b decreased both Mcl-1 expression and the IC50 to sorafenib. Conclusions: Cellular expression of full-length HCV increases sensitivity to sorafenib by the miRNA-dependent modulation of Mcl-1 and apoptosis. Modulation of miRNA responses may be a useful strategy to enhance response to chemotherapy in HCC.

Original languageEnglish (US)
Pages (from-to)957-966
Number of pages10
JournalClinical Cancer Research
Volume16
Issue number3
DOIs
StatePublished - Feb 1 2010
Externally publishedYes

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MicroRNAs
Hepacivirus
Hepatocytes
Liver Neoplasms
Proteins
Viral Proteins
Inhibitory Concentration 50
Caspase 7
Drug Therapy
Poly(ADP-ribose) Polymerases
Chronic Hepatitis C
Virus Diseases
Caspases
Caspase 3
Transfection
Real-Time Polymerase Chain Reaction
Research Design
Down-Regulation
Western Blotting
Genome

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Hepatitis C virus proteins modulate microRNA expression and chemosensitivity in malignant hepatocytes. / Braconi, Chiara; Valeri, Nicola; Gasparini, Pierluigi; Huang, Nianyuan; Taccioli, Cristian; Nuovo, Gerard; Suzuki, Tetsuro; Croce, Carlo Maria; Patel, Tushar C.

In: Clinical Cancer Research, Vol. 16, No. 3, 01.02.2010, p. 957-966.

Research output: Contribution to journalArticle

Braconi, C, Valeri, N, Gasparini, P, Huang, N, Taccioli, C, Nuovo, G, Suzuki, T, Croce, CM & Patel, TC 2010, 'Hepatitis C virus proteins modulate microRNA expression and chemosensitivity in malignant hepatocytes', Clinical Cancer Research, vol. 16, no. 3, pp. 957-966. https://doi.org/10.1158/1078-0432.CCR-09-2123
Braconi, Chiara ; Valeri, Nicola ; Gasparini, Pierluigi ; Huang, Nianyuan ; Taccioli, Cristian ; Nuovo, Gerard ; Suzuki, Tetsuro ; Croce, Carlo Maria ; Patel, Tushar C. / Hepatitis C virus proteins modulate microRNA expression and chemosensitivity in malignant hepatocytes. In: Clinical Cancer Research. 2010 ; Vol. 16, No. 3. pp. 957-966.
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T1 - Hepatitis C virus proteins modulate microRNA expression and chemosensitivity in malignant hepatocytes

AU - Braconi, Chiara

AU - Valeri, Nicola

AU - Gasparini, Pierluigi

AU - Huang, Nianyuan

AU - Taccioli, Cristian

AU - Nuovo, Gerard

AU - Suzuki, Tetsuro

AU - Croce, Carlo Maria

AU - Patel, Tushar C

PY - 2010/2/1

Y1 - 2010/2/1

N2 - Purpose: Hepatocellular cancer (HCC) is highly resistant to chemotherapy and is associated with poor prognosis. Chronic hepatitis C virus (HCV) infection is a major cause of HCC. However, the effect of viral proteins in mediating chemosensitivity in tumor cells is unknown. We postulated that HCV viral proteins could modulate therapeutic responses by altering host cell microRNA (miRNA) expression. Experimental Design: HepG2 malignant hepatocytes were stably transfected with full-length HCV genome (Hep-394) or an empty vector (Hep-SWX). MiRNA profiling was done by using a custom microarray, and the expression of selected miRNAs was validated by real-time PCR. Protein expression was assessed by Western blotting, whereas caspase activation was assessed by a luminometric assay. Results: The IC50 to sorafenib was lower in Hep-394 compared with Hep-SWX control cells. Alterations in miRNA expression occurred with 10 miRNAs downregulated >2-fold and 23 miRNAs upregulated >2-fold in Hep-394 cells compared with controls. Of these, miR-193b was overexpressed by 5-fold in Hep-394 cells. miR-193b was predicted to target Mcl-1, an antiapoptotic protein that can modulate the response to sorafenib. The expression of Mcl-1 was decreased, and basal caspase-3/7 activity and poly ADP ribose polymerase cleavage were increased in Hep-394 cells compared with controls. Moreover, transfection with precursors to miR-193b decreased both Mcl-1 expression and the IC50 to sorafenib. Conclusions: Cellular expression of full-length HCV increases sensitivity to sorafenib by the miRNA-dependent modulation of Mcl-1 and apoptosis. Modulation of miRNA responses may be a useful strategy to enhance response to chemotherapy in HCC.

AB - Purpose: Hepatocellular cancer (HCC) is highly resistant to chemotherapy and is associated with poor prognosis. Chronic hepatitis C virus (HCV) infection is a major cause of HCC. However, the effect of viral proteins in mediating chemosensitivity in tumor cells is unknown. We postulated that HCV viral proteins could modulate therapeutic responses by altering host cell microRNA (miRNA) expression. Experimental Design: HepG2 malignant hepatocytes were stably transfected with full-length HCV genome (Hep-394) or an empty vector (Hep-SWX). MiRNA profiling was done by using a custom microarray, and the expression of selected miRNAs was validated by real-time PCR. Protein expression was assessed by Western blotting, whereas caspase activation was assessed by a luminometric assay. Results: The IC50 to sorafenib was lower in Hep-394 compared with Hep-SWX control cells. Alterations in miRNA expression occurred with 10 miRNAs downregulated >2-fold and 23 miRNAs upregulated >2-fold in Hep-394 cells compared with controls. Of these, miR-193b was overexpressed by 5-fold in Hep-394 cells. miR-193b was predicted to target Mcl-1, an antiapoptotic protein that can modulate the response to sorafenib. The expression of Mcl-1 was decreased, and basal caspase-3/7 activity and poly ADP ribose polymerase cleavage were increased in Hep-394 cells compared with controls. Moreover, transfection with precursors to miR-193b decreased both Mcl-1 expression and the IC50 to sorafenib. Conclusions: Cellular expression of full-length HCV increases sensitivity to sorafenib by the miRNA-dependent modulation of Mcl-1 and apoptosis. Modulation of miRNA responses may be a useful strategy to enhance response to chemotherapy in HCC.

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