Survival pathway of cholangiocarcinoma via 
AKT/mTOR signaling to escape RAF/MEK/ERK
pathway inhibition by sorafenib

Yokoi,Kenta; Kobayashi,Akira; Motoyama,Hiroaki; Kitazawa,Masato; Shimizu,Akira; Notake,Tsuyoshi; Yokoyama,Takahide; Matsumura,Tomio; Takeoka,Michiko; Miyagawa,Shin-Ichi

doi:10.3892/or.2017.6153

Survival pathway of cholangiocarcinoma via AKT/mTOR signaling to escape RAF/MEK/ERK pathway inhibition by sorafenib

Authors:
- Kenta Yokoi
- Akira Kobayashi
- Hiroaki Motoyama
- Masato Kitazawa
- Akira Shimizu
- Tsuyoshi Notake
- Takahide Yokoyama
- Tomio Matsumura
- Michiko Takeoka
- Shin-Ichi Miyagawa
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Affiliations: Department of Surgery, Shinshu University School of Medicine, Matsumoto 390-8621, Japan, Department of Molecular Oncology, Shinshu University Graduate School of Medicine, Matsumoto 390-8621, Japan
Published online on: December 13, 2017 https://doi.org/10.3892/or.2017.6153
Pages: 843-850

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Abstract

Cholangiocarcinoma (CCC) is a strongly aggressive malignancy for which surgical resection is the only potential curative therapy. Sorafenib, a multikinase inhibitor of the RAF/MEK/ERK pathway, is a molecular-targeted drug that is approved for hepatocellular carcinoma (HCC) but not for CCC. The differences in signaling pathway characteristics under sorafenib treatment between HCC (HLF, Huh7, PLC/PRF/5) and CCC (RBE, YSCCC, Huh28) cell lines were therefore investigated using cell proliferation, western blotting, and apoptosis analyses. Sorafenib inhibited cell growth significantly less in CCC cells than in HCC cells, with lower suppression of ERK phosphorylation. Significantly decreased AKT Ser473 phosphorylation in HCC cells, and conversely enhanced phosphorylation of AKT Ser473 and mTORC2 in CCC cells, were observed with sorafenib treatment. Disassembly of the mTORC2 complex in RBE cells with siRNA targeting Rictor resulted in the downregulation of AKT Ser473 phosphorylation and enhanced apoptosis presumably via increased FOXO1, which consequently suppressed RBE cell proliferation. Phosphorylation of mTORC1 and autophagy were not influenced by sorafenib in CCC cells. Simultaneous administration of everolimus to suppress activated mTORC1 in RBE cells revealed that combined everolimus and sorafenib treatment under mTORC2 disassembly could enhance growth inhibition through the suppression of both sorafenib- and everolimus-dependent AKT Ser473 phosphorylation in addition to the inhibition of mTORC1 phosphorylation. Prevention of escape by AKT/mTOR signaling from the RAF/MEK/ERK pathway in sorafenib treatment by suppressing mTORC2 activity may lead to promising new approaches in CCC therapy.

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