Regulation of interferon signaling and HCV‑RNA replication by extracellular matrix

Takuya Kuwashiro, Shinji Iwane, Xia Jinghe, Sachiko Matsuhashi, Yuichiro Eguchi, Keizo Anzai, Kazuma Fujimoto, Toshihiko Mizuta, Naoya Sakamoto, Masanori Ikeda, Nobuyuki Kato, Iwata Ozaki

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Although interferon (IFN)‑based treatment of patients with chronic hepatitis C virus (HCV) infection is widely applied, treatment resistance is often observed in patients with advanced liver fibrosis. Given that the molecular mechanisms of IFN resistance in liver fibrosis remain elusive, the present study investigated the effects of extracellular matrix (ECM) on IFN signaling in hepatic cells. The native HuH‑7 human hepatoma cell line and HuH‑7 cells were stably transfected with full‑length HCV‑RNA fused with Renilla luciferase (OR6 cells) were cultured on ECM‑coated dishes or non‑coated plastic dishes (NDs), and treated with human IFN‑α. In Huh‑7 cells cultured on coated dishes, the IFN‑stimulated response element (ISRE) luciferase activity was measured following ISRE plasmid transfection and the expression of IFN‑stimulated genes (ISG) were significantly lower than those in cells cultured on NDs. In addition, after IFN‑α treatment, the amount of HCV‑RNA and viral protein produced by OR6 cells cultured on coated dishes was higher than that produced by cells cultured on NDs. When cells were treated with β1‑integrin‑blocking antibody to disrupt the cell‑matrix interaction, the ISRE luciferase activity was restored, and the protein expression of ISG was increased, while that of HCV proteins was suppressed. Treatment of cells with integrin‑linked kinase (ILK) inhibitor or focal adhesion kinase (FAK) inhibitor restored the ISRE luciferase activity and expression of ISG proteins. These results suggested that β1‑integrin‑mediated signals affected the IFN signaling and promoted HCV replication. Therefore, the accumulation of ECM in liver fibrosis may impair IFN signaling through β1‑integrin‑mediated signaling involving ILK and FAK.

Original languageEnglish
Pages (from-to)957-965
Number of pages9
JournalInternational Journal of Molecular Medicine
Volume42
Issue number2
DOIs
Publication statusPublished - Aug 1 2018

Fingerprint

Interferons
Extracellular Matrix
Response Elements
Cultured Cells
Luciferases
Liver Cirrhosis
Plastics
Focal Adhesion Protein-Tyrosine Kinases
Phosphotransferases
Renilla Luciferases
Gene Expression
Proteins
Viral Proteins
Virus Diseases
Therapeutics
Virus Replication
Transfection
Hepatocytes
Hepatocellular Carcinoma
Plasmids

ASJC Scopus subject areas

  • Genetics

Cite this

Kuwashiro, T., Iwane, S., Jinghe, X., Matsuhashi, S., Eguchi, Y., Anzai, K., ... Ozaki, I. (2018). Regulation of interferon signaling and HCV‑RNA replication by extracellular matrix. International Journal of Molecular Medicine, 42(2), 957-965. https://doi.org/10.3892/ijmm.2018.3693

Regulation of interferon signaling and HCV‑RNA replication by extracellular matrix. / Kuwashiro, Takuya; Iwane, Shinji; Jinghe, Xia; Matsuhashi, Sachiko; Eguchi, Yuichiro; Anzai, Keizo; Fujimoto, Kazuma; Mizuta, Toshihiko; Sakamoto, Naoya; Ikeda, Masanori; Kato, Nobuyuki; Ozaki, Iwata.

In: International Journal of Molecular Medicine, Vol. 42, No. 2, 01.08.2018, p. 957-965.

Research output: Contribution to journalArticle

Kuwashiro, T, Iwane, S, Jinghe, X, Matsuhashi, S, Eguchi, Y, Anzai, K, Fujimoto, K, Mizuta, T, Sakamoto, N, Ikeda, M, Kato, N & Ozaki, I 2018, 'Regulation of interferon signaling and HCV‑RNA replication by extracellular matrix', International Journal of Molecular Medicine, vol. 42, no. 2, pp. 957-965. https://doi.org/10.3892/ijmm.2018.3693
Kuwashiro, Takuya ; Iwane, Shinji ; Jinghe, Xia ; Matsuhashi, Sachiko ; Eguchi, Yuichiro ; Anzai, Keizo ; Fujimoto, Kazuma ; Mizuta, Toshihiko ; Sakamoto, Naoya ; Ikeda, Masanori ; Kato, Nobuyuki ; Ozaki, Iwata. / Regulation of interferon signaling and HCV‑RNA replication by extracellular matrix. In: International Journal of Molecular Medicine. 2018 ; Vol. 42, No. 2. pp. 957-965.
@article{b97342cd07de44e291eac952ed7bd404,
title = "Regulation of interferon signaling and HCV‑RNA replication by extracellular matrix",
abstract = "Although interferon (IFN)‑based treatment of patients with chronic hepatitis C virus (HCV) infection is widely applied, treatment resistance is often observed in patients with advanced liver fibrosis. Given that the molecular mechanisms of IFN resistance in liver fibrosis remain elusive, the present study investigated the effects of extracellular matrix (ECM) on IFN signaling in hepatic cells. The native HuH‑7 human hepatoma cell line and HuH‑7 cells were stably transfected with full‑length HCV‑RNA fused with Renilla luciferase (OR6 cells) were cultured on ECM‑coated dishes or non‑coated plastic dishes (NDs), and treated with human IFN‑α. In Huh‑7 cells cultured on coated dishes, the IFN‑stimulated response element (ISRE) luciferase activity was measured following ISRE plasmid transfection and the expression of IFN‑stimulated genes (ISG) were significantly lower than those in cells cultured on NDs. In addition, after IFN‑α treatment, the amount of HCV‑RNA and viral protein produced by OR6 cells cultured on coated dishes was higher than that produced by cells cultured on NDs. When cells were treated with β1‑integrin‑blocking antibody to disrupt the cell‑matrix interaction, the ISRE luciferase activity was restored, and the protein expression of ISG was increased, while that of HCV proteins was suppressed. Treatment of cells with integrin‑linked kinase (ILK) inhibitor or focal adhesion kinase (FAK) inhibitor restored the ISRE luciferase activity and expression of ISG proteins. These results suggested that β1‑integrin‑mediated signals affected the IFN signaling and promoted HCV replication. Therefore, the accumulation of ECM in liver fibrosis may impair IFN signaling through β1‑integrin‑mediated signaling involving ILK and FAK.",
author = "Takuya Kuwashiro and Shinji Iwane and Xia Jinghe and Sachiko Matsuhashi and Yuichiro Eguchi and Keizo Anzai and Kazuma Fujimoto and Toshihiko Mizuta and Naoya Sakamoto and Masanori Ikeda and Nobuyuki Kato and Iwata Ozaki",
year = "2018",
month = "8",
day = "1",
doi = "10.3892/ijmm.2018.3693",
language = "English",
volume = "42",
pages = "957--965",
journal = "International Journal of Molecular Medicine",
issn = "1107-3756",
publisher = "Spandidos Publications",
number = "2",

}

TY - JOUR

T1 - Regulation of interferon signaling and HCV‑RNA replication by extracellular matrix

AU - Kuwashiro, Takuya

AU - Iwane, Shinji

AU - Jinghe, Xia

AU - Matsuhashi, Sachiko

AU - Eguchi, Yuichiro

AU - Anzai, Keizo

AU - Fujimoto, Kazuma

AU - Mizuta, Toshihiko

AU - Sakamoto, Naoya

AU - Ikeda, Masanori

AU - Kato, Nobuyuki

AU - Ozaki, Iwata

PY - 2018/8/1

Y1 - 2018/8/1

N2 - Although interferon (IFN)‑based treatment of patients with chronic hepatitis C virus (HCV) infection is widely applied, treatment resistance is often observed in patients with advanced liver fibrosis. Given that the molecular mechanisms of IFN resistance in liver fibrosis remain elusive, the present study investigated the effects of extracellular matrix (ECM) on IFN signaling in hepatic cells. The native HuH‑7 human hepatoma cell line and HuH‑7 cells were stably transfected with full‑length HCV‑RNA fused with Renilla luciferase (OR6 cells) were cultured on ECM‑coated dishes or non‑coated plastic dishes (NDs), and treated with human IFN‑α. In Huh‑7 cells cultured on coated dishes, the IFN‑stimulated response element (ISRE) luciferase activity was measured following ISRE plasmid transfection and the expression of IFN‑stimulated genes (ISG) were significantly lower than those in cells cultured on NDs. In addition, after IFN‑α treatment, the amount of HCV‑RNA and viral protein produced by OR6 cells cultured on coated dishes was higher than that produced by cells cultured on NDs. When cells were treated with β1‑integrin‑blocking antibody to disrupt the cell‑matrix interaction, the ISRE luciferase activity was restored, and the protein expression of ISG was increased, while that of HCV proteins was suppressed. Treatment of cells with integrin‑linked kinase (ILK) inhibitor or focal adhesion kinase (FAK) inhibitor restored the ISRE luciferase activity and expression of ISG proteins. These results suggested that β1‑integrin‑mediated signals affected the IFN signaling and promoted HCV replication. Therefore, the accumulation of ECM in liver fibrosis may impair IFN signaling through β1‑integrin‑mediated signaling involving ILK and FAK.

AB - Although interferon (IFN)‑based treatment of patients with chronic hepatitis C virus (HCV) infection is widely applied, treatment resistance is often observed in patients with advanced liver fibrosis. Given that the molecular mechanisms of IFN resistance in liver fibrosis remain elusive, the present study investigated the effects of extracellular matrix (ECM) on IFN signaling in hepatic cells. The native HuH‑7 human hepatoma cell line and HuH‑7 cells were stably transfected with full‑length HCV‑RNA fused with Renilla luciferase (OR6 cells) were cultured on ECM‑coated dishes or non‑coated plastic dishes (NDs), and treated with human IFN‑α. In Huh‑7 cells cultured on coated dishes, the IFN‑stimulated response element (ISRE) luciferase activity was measured following ISRE plasmid transfection and the expression of IFN‑stimulated genes (ISG) were significantly lower than those in cells cultured on NDs. In addition, after IFN‑α treatment, the amount of HCV‑RNA and viral protein produced by OR6 cells cultured on coated dishes was higher than that produced by cells cultured on NDs. When cells were treated with β1‑integrin‑blocking antibody to disrupt the cell‑matrix interaction, the ISRE luciferase activity was restored, and the protein expression of ISG was increased, while that of HCV proteins was suppressed. Treatment of cells with integrin‑linked kinase (ILK) inhibitor or focal adhesion kinase (FAK) inhibitor restored the ISRE luciferase activity and expression of ISG proteins. These results suggested that β1‑integrin‑mediated signals affected the IFN signaling and promoted HCV replication. Therefore, the accumulation of ECM in liver fibrosis may impair IFN signaling through β1‑integrin‑mediated signaling involving ILK and FAK.

UR - http://www.scopus.com/inward/record.url?scp=85054777378&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85054777378&partnerID=8YFLogxK

U2 - 10.3892/ijmm.2018.3693

DO - 10.3892/ijmm.2018.3693

M3 - Article

C2 - 29786754

AN - SCOPUS:85054777378

VL - 42

SP - 957

EP - 965

JO - International Journal of Molecular Medicine

JF - International Journal of Molecular Medicine

SN - 1107-3756

IS - 2

ER -