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Sorafenib enhances proteasome inhibitor-induced cell death via inactivation of Akt and stress-activated protein kinases

  • Original Article—Liver, Pancreas, and Biliary Tract
  • Published:
Journal of Gastroenterology Aims and scope Submit manuscript

Abstract

Background

Advanced hepatocellular carcinoma (HCC) responds poorly to conventional systemic therapies. Therefore, new effective therapy strategies are urgently needed. Molecular targeted therapies have entered the field of anti-neoplastic treatment and are being used on their own and in combination with other drugs. Sorafenib inhibits proliferation and angiogenesis of HCC by suppressing the Raf serine/threonine kinases and the receptor tyrosine kinases. The proteasome inhibitor bortezomib has shown activity in a variety of solid tumors, including HCC. However, the precise anti-proliferative mechanisms of these agents remain unclear.

Methods

We treated human hepatoma cell lines (Huh7 and Hep3B) and immortalized human hepatocyte (OUMS29) with sorafenib and/or proteasome inhibitors, including epoxomicin and acetyl-leucyl-leucyl-norleucinal. Cytotoxic effects were examined by morphometric analyses of apoptosis and necrosis. Apoptosis was also evaluated by Western blotting of keratin18, PARP and caspase3. The activity of Akt and stress-activated protein kinases was examined by Western blotting.

Results

Both sorafenib and proteasome inhibitors induced apoptosis in Huh7 and OUMS29. However, sorafenib attenuated proteasome inhibitor-induced apoptosis. Sorafenib induced necrosis, especially in combination with proteasome inhibitors. Sorafenib induced down-regulation of Akt synergistically in combination with proteasome inhibitors in Huh7. Sorafenib inhibited both the JNK and p38 pathways in a time- and dose-dependent manner. In addition, sorafenib also inhibited proteasome inhibitor-mediated JNK and p38 activation in both Huh7 and OUMS29.

Conclusions

Sorafenib enhances the anti-proliferative effect of proteasome inhibitors in part by inactivating the Akt signaling pathway and modulating stress-activated protein kinases. The combination of these agents could be an ideal molecular targeted therapy for HCC.

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Abbreviations

Ab:

Antibody

ALLN:

Acetyl-leucyl-leucyl-norleucinal

CREB:

cAMP response element-binding protein

DAPI:

4′6-diamidino-2-phenylindole

ER:

Endoplasmic reticulum

ERK:

Extracellular signal-regulated protein kinase

HCC:

Hepatocellular carcinoma

IGF-1:

Insulin-like growth factor-1

JNK:

c-Jun N-terminal kinase

K:

Keratin

MAPK:

Mitogen-activated protein kinase

MEK:

MAPK/ERK kinase

mTOR:

Mammalian target of rapamycin

mTORC:

Mammalian target of rapamycin complex

NFκB:

Nuclear factor-kappa B

PARP:

Poly-ADP-ribose-polymerase

PDGFR:

Platelet-derived growth factor receptor

PI:

Proteasome inhibitor

PI3K:

Phosphoinositide 3-kinase

ROS:

Reactive oxygen species

TNF-α:

Tumor necrosis factor-α

TRAF2:

Tumor necrosis factor receptor-associated factor-2

VEGFR:

Vascular endothelial growth factor receptor

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Acknowledgments

Our work is supported by in part by a grant-in-aid (2059798, 23591000) from the Ministry of Education, Culture, Sports, Science and Technology, Japan and UOEH Grant for Advanced Research (H22-1) to M.H. We thank Ms. Y. Katsuki and Ms. H. Mihara for their expert technical assistance.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Third Department of Internal Medicine, School of Medicine, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu, 807-8555, Japan

    Yuichi Honma & Masaru Harada

  2. Department of Gastroenterology and Hepatology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan

    Satoshi Shimizu & Tetsuo Takehara

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  1. Yuichi Honma
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  2. Satoshi Shimizu
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  3. Tetsuo Takehara
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  4. Masaru Harada
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Correspondence to Masaru Harada.

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Honma, Y., Shimizu, S., Takehara, T. et al. Sorafenib enhances proteasome inhibitor-induced cell death via inactivation of Akt and stress-activated protein kinases. J Gastroenterol 49, 517–526 (2014). https://doi.org/10.1007/s00535-013-0796-z

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  • DOI: https://doi.org/10.1007/s00535-013-0796-z

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