title: 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.
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.
authors.affiliation: Department of Medical Oncology, Graduate School of Medicine, Chiba University, Inohana, Chuo-ku, Chiba 260-8670, Japan.
authors.forename: Ryota
authors.initials: R
authors.lastname: Kurimoto
authors.affiliation: Department of Medical Oncology, Graduate School of Medicine, Chiba University, Inohana, Chuo-ku, Chiba 260-8670, Japan.
authors.forename: Shunichiro
authors.initials: S
authors.lastname: Iwasawa
authors.affiliation: Department of Medical Oncology, Graduate School of Medicine, Chiba University, Inohana, Chuo-ku, Chiba 260-8670, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Ebata
authors.affiliation: Department of Respirology, Graduate School of Medicine, Chiba University, Inohana, Chuo-ku, Chiba 260-8670, Japan.
authors.forename: Tsukasa
authors.initials: T
authors.lastname: Ishiwata
authors.affiliation: Department of Medical Oncology, Faculty of Medicine, University of Tsukuba, Tennodai, Tsukuba 305-8575, Japan.
authors.forename: Ikuo
authors.initials: I
authors.lastname: Sekine
authors.affiliation: Department of Respirology, Graduate School of Medicine, Chiba University, Inohana, Chuo-ku, Chiba 260-8670, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Tada
authors.affiliation: Department of Respirology, Graduate School of Medicine, Chiba University, Inohana, Chuo-ku, Chiba 260-8670, Japan.
authors.forename: Koichiro
authors.initials: K
authors.lastname: Tatsumi
authors.affiliation: Department of Cellular and Molecular Medicine, Graduate School of Medicine, Chiba University, Inohana, Chuo-ku, Chiba 260-8670, Japan.
authors.forename: Shuhei
authors.initials: S
authors.lastname: Koide
authors.affiliation: Department of Cellular and Molecular Medicine, Graduate School of Medicine, Chiba University, Inohana, Chuo-ku, Chiba 260-8670, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Iwama
authors.affiliation: Department of Medical Oncology, Graduate School of Medicine, Chiba University, Inohana, Chuo-ku, Chiba 260-8670, Japan.
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Takiguchi
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics* / Q000378:metabolism; D000077192:Adenocarcinoma of Lung; D000970:Antineoplastic Agents / Q000494:pharmacology; D060890:B7-H1 Antigen / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D019008:Drug Resistance, Neoplasm / Q000187:drug effects*; D058750:Epithelial-Mesenchymal Transition; D066246:ErbB Receptors / Q000235:genetics*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D000077156:Gefitinib; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007211:Indoles / Q000494:pharmacology*; D008175:Lung Neoplasms / Q000235:genetics* / Q000378:metabolism; D008687:Metformin / Q000494:pharmacology*; D009154:Mutation; D011687:Purines / Q000494:pharmacology*; D011799:Quinazolines / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000494:pharmacology
chemical_list: D000970:Antineoplastic Agents; D060890:B7-H1 Antigen; C423236:CD274 protein, human; D007211:Indoles; D011687:Purines; D011799:Quinazolines; D016212:Transforming Growth Factor beta; D016222:Fibroblast Growth Factor 2; D008687:Metformin; C512478:EGFR protein, human; D066246:ErbB Receptors; C572919:PP242; D002945:Cisplatin; D000077156:Gefitinib; D004121:Dimethyl Sulfoxide