Drug resistance originating from a TGF-β/FGF-2-driven epithelial-to-mesenchymal transition and its reversion in human lung adenocarcinoma cell lines harboring an EGFR mutation

Kurimoto,Ryota; Iwasawa,Shunichiro; Ebata,Takahiro; Ishiwata,Tsukasa; Sekine,Ikuo; Tada,Yuji; Tatsumi,Koichiro; Koide,Shuhei; Iwama,Atsushi; Takiguchi,Yuichi

doi:10.3892/ijo.2016.3419

Drug resistance originating from a TGF-β/FGF-2-driven epithelial-to-mesenchymal transition and its reversion in human lung adenocarcinoma cell lines harboring an EGFR mutation

Authors:
- Ryota Kurimoto
- Shunichiro Iwasawa
- Takahiro Ebata
- Tsukasa Ishiwata
- Ikuo Sekine
- Yuji Tada
- Koichiro Tatsumi
- Shuhei Koide
- Atsushi Iwama
- Yuichi Takiguchi
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Affiliations: Department of Medical Oncology, Graduate School of Medicine, Chiba University, Inohana, Chuo-ku, Chiba 260-8670, Japan, Department of Respirology, Graduate School of Medicine, Chiba University, Inohana, Chuo-ku, Chiba 260-8670, Japan, Department of Medical Oncology, Faculty of Medicine, University of Tsukuba, Tennodai, Tsukuba 305-8575, Japan, Department of Cellular and Molecular Medicine, Graduate School of Medicine, Chiba University, Inohana, Chuo-ku, Chiba 260-8670, Japan
Published online on: March 4, 2016 https://doi.org/10.3892/ijo.2016.3419
Pages: 1825-1836
Copyright: © Kurimoto et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Epithelial-to-mesenchymal transition (EMT) is a malignant cancer phenotype characterized by augmented invasion and metastasis, chemoresistance, and escape from host-immunity. This study sought to identify efficient methods for inducing EMT reversion, to evaluate alterations in chemosensitivity and immune-protectiveness, and to elucidate the underlying mechanisms. In this study, the human lung adenocarcinoma cell lines PC-9 and HCC-827, harboring an EGFR mutation, were treated with TGF-β and FGF-2 to induce EMT. The phenotypic alterations were evaluated by RT-PCR, fluorescent immunohistochemistry, cell-mobility, and flow cytometry. Chemosensitivity to gefitinib and cisplatin was evaluated using an MTT assay and apoptosis. Immune-protectiveness was evaluated by PD-L1 expression. A combination of TGF-β and FGF-2 efficiently induced EMT in both cell lines: through Smad3 pathway in PC-9, and through Smad3, MEK/Erk, and mTOR pathways in HCC-827. The mTOR inhibitor PP242, metformin, and DMSO reverted EMT to different extent and through different pathways, depending on the cell lines. EMT induction reduced the sensitivity to gefitinib in both cell lines and to cisplatin in HCC-827, and it increased PD-L1 expression in both cell lines. EMT reversion using each of the 3 agents partly restored chemosensitivity and suppressed PD-L1 expression. Thus, chemoresistance and increased PD-L1 expression caused by EMT can be successfully reverted by EMT-reverting agents.

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