CD44 variant–dependent regulation of redox balance in EGFR mutation–positive non–small cell lung cancer: A target for treatment

Yuko Kawano, Eiji Iwama, Kenji Tsuchihashi, Daisuke Shibahara, Taishi Harada, Kentaro Tanaka, Osamu Nagano, Hideyuki Saya, Yoichi Nakanishi, Isamu Okamoto

研究成果: ジャーナルへの寄稿記事

4 引用 (Scopus)

抄録

Objectives The regulation of redox balance in cancer cells is an important factor in tumor development and chemoresistance, with oncogene activation having been shown to induce the generation of reactive oxygen species (ROS). Activating mutations of the epidermal growth factor receptor gene (EGFR) are oncogenic drivers in non–small cell lung cancer (NSCLC), but it has remained unknown whether ligand-independent EGFR signaling conferred by EGFR mutation triggers ROS generation in NSCLC cells. Materials and Methods HEK293T cells were transfected with an expression vector for mutant EGFR. The expression of CD44 variant (CD44v) isoforms in NSCLC cell lines was evaluated by flow cytometry. Cells were depleted of CD44v by RNA interference and assayed for ROS and glutathione (GSH) levels. The effect of CD44v on cisplatin sensitivity was evaluated in vitro with the MTS assay. Results EGFR signaling due to EGFR mutation increased ROS levels in transfected HEK293T cells. The expression of CD44v isoforms was found to be inversely correlated with basal ROS levels in EGFR mutation–positive NSCLC cell lines. Knockdown of CD44v induced depletion of intracellular GSH and increased ROS levels in EGFR-mutated NSCLC cells that express CD44v at a high level (CD44vhigh). In addition, depletion of GSH by treatment with buthionine-[S, R]-sulfoximine induced marked accumulation of ROS and enhanced the cytotoxicity of cisplatin in CD44vhigh EGFR-mutated NSCLC cells but not in corresponding CD44vlow cells. This enhancement of cisplatin cytotoxicity by GSH depletion was prevented by treatment with the antioxidant N-acetyl-L-cysteine. Knockdown of CD44v also enhanced cisplatin cytotoxicity in CD44vhigh EGFR mutation–positive NSCLC cells but not in CD44vlow cells. Conclusion Our results thus implicate CD44v in redox adaptation and as a potential target for treatment in CD44vhigh EGFR-mutated NSCLC cells.

元の言語英語
ページ(範囲)72-78
ページ数7
ジャーナルLung Cancer
113
DOI
出版物ステータス出版済み - 11 1 2017

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erbB-1 Genes
Non-Small Cell Lung Carcinoma
Oxidation-Reduction
Reactive Oxygen Species
Cisplatin
Mutation
Protein Isoforms
Cell Line
Acetylcysteine
RNA Interference
Oncogenes
Glutathione
Neoplasms
Flow Cytometry
Antioxidants

All Science Journal Classification (ASJC) codes

  • Oncology
  • Pulmonary and Respiratory Medicine
  • Cancer Research

これを引用

CD44 variant–dependent regulation of redox balance in EGFR mutation–positive non–small cell lung cancer : A target for treatment. / Kawano, Yuko; Iwama, Eiji; Tsuchihashi, Kenji; Shibahara, Daisuke; Harada, Taishi; Tanaka, Kentaro; Nagano, Osamu; Saya, Hideyuki; Nakanishi, Yoichi; Okamoto, Isamu.

:: Lung Cancer, 巻 113, 01.11.2017, p. 72-78.

研究成果: ジャーナルへの寄稿記事

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title = "CD44 variant–dependent regulation of redox balance in EGFR mutation–positive non–small cell lung cancer: A target for treatment",
abstract = "Objectives The regulation of redox balance in cancer cells is an important factor in tumor development and chemoresistance, with oncogene activation having been shown to induce the generation of reactive oxygen species (ROS). Activating mutations of the epidermal growth factor receptor gene (EGFR) are oncogenic drivers in non–small cell lung cancer (NSCLC), but it has remained unknown whether ligand-independent EGFR signaling conferred by EGFR mutation triggers ROS generation in NSCLC cells. Materials and Methods HEK293T cells were transfected with an expression vector for mutant EGFR. The expression of CD44 variant (CD44v) isoforms in NSCLC cell lines was evaluated by flow cytometry. Cells were depleted of CD44v by RNA interference and assayed for ROS and glutathione (GSH) levels. The effect of CD44v on cisplatin sensitivity was evaluated in vitro with the MTS assay. Results EGFR signaling due to EGFR mutation increased ROS levels in transfected HEK293T cells. The expression of CD44v isoforms was found to be inversely correlated with basal ROS levels in EGFR mutation–positive NSCLC cell lines. Knockdown of CD44v induced depletion of intracellular GSH and increased ROS levels in EGFR-mutated NSCLC cells that express CD44v at a high level (CD44vhigh). In addition, depletion of GSH by treatment with buthionine-[S, R]-sulfoximine induced marked accumulation of ROS and enhanced the cytotoxicity of cisplatin in CD44vhigh EGFR-mutated NSCLC cells but not in corresponding CD44vlow cells. This enhancement of cisplatin cytotoxicity by GSH depletion was prevented by treatment with the antioxidant N-acetyl-L-cysteine. Knockdown of CD44v also enhanced cisplatin cytotoxicity in CD44vhigh EGFR mutation–positive NSCLC cells but not in CD44vlow cells. Conclusion Our results thus implicate CD44v in redox adaptation and as a potential target for treatment in CD44vhigh EGFR-mutated NSCLC cells.",
author = "Yuko Kawano and Eiji Iwama and Kenji Tsuchihashi and Daisuke Shibahara and Taishi Harada and Kentaro Tanaka and Osamu Nagano and Hideyuki Saya and Yoichi Nakanishi and Isamu Okamoto",
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T1 - CD44 variant–dependent regulation of redox balance in EGFR mutation–positive non–small cell lung cancer

T2 - A target for treatment

AU - Kawano, Yuko

AU - Iwama, Eiji

AU - Tsuchihashi, Kenji

AU - Shibahara, Daisuke

AU - Harada, Taishi

AU - Tanaka, Kentaro

AU - Nagano, Osamu

AU - Saya, Hideyuki

AU - Nakanishi, Yoichi

AU - Okamoto, Isamu

PY - 2017/11/1

Y1 - 2017/11/1

N2 - Objectives The regulation of redox balance in cancer cells is an important factor in tumor development and chemoresistance, with oncogene activation having been shown to induce the generation of reactive oxygen species (ROS). Activating mutations of the epidermal growth factor receptor gene (EGFR) are oncogenic drivers in non–small cell lung cancer (NSCLC), but it has remained unknown whether ligand-independent EGFR signaling conferred by EGFR mutation triggers ROS generation in NSCLC cells. Materials and Methods HEK293T cells were transfected with an expression vector for mutant EGFR. The expression of CD44 variant (CD44v) isoforms in NSCLC cell lines was evaluated by flow cytometry. Cells were depleted of CD44v by RNA interference and assayed for ROS and glutathione (GSH) levels. The effect of CD44v on cisplatin sensitivity was evaluated in vitro with the MTS assay. Results EGFR signaling due to EGFR mutation increased ROS levels in transfected HEK293T cells. The expression of CD44v isoforms was found to be inversely correlated with basal ROS levels in EGFR mutation–positive NSCLC cell lines. Knockdown of CD44v induced depletion of intracellular GSH and increased ROS levels in EGFR-mutated NSCLC cells that express CD44v at a high level (CD44vhigh). In addition, depletion of GSH by treatment with buthionine-[S, R]-sulfoximine induced marked accumulation of ROS and enhanced the cytotoxicity of cisplatin in CD44vhigh EGFR-mutated NSCLC cells but not in corresponding CD44vlow cells. This enhancement of cisplatin cytotoxicity by GSH depletion was prevented by treatment with the antioxidant N-acetyl-L-cysteine. Knockdown of CD44v also enhanced cisplatin cytotoxicity in CD44vhigh EGFR mutation–positive NSCLC cells but not in CD44vlow cells. Conclusion Our results thus implicate CD44v in redox adaptation and as a potential target for treatment in CD44vhigh EGFR-mutated NSCLC cells.

AB - Objectives The regulation of redox balance in cancer cells is an important factor in tumor development and chemoresistance, with oncogene activation having been shown to induce the generation of reactive oxygen species (ROS). Activating mutations of the epidermal growth factor receptor gene (EGFR) are oncogenic drivers in non–small cell lung cancer (NSCLC), but it has remained unknown whether ligand-independent EGFR signaling conferred by EGFR mutation triggers ROS generation in NSCLC cells. Materials and Methods HEK293T cells were transfected with an expression vector for mutant EGFR. The expression of CD44 variant (CD44v) isoforms in NSCLC cell lines was evaluated by flow cytometry. Cells were depleted of CD44v by RNA interference and assayed for ROS and glutathione (GSH) levels. The effect of CD44v on cisplatin sensitivity was evaluated in vitro with the MTS assay. Results EGFR signaling due to EGFR mutation increased ROS levels in transfected HEK293T cells. The expression of CD44v isoforms was found to be inversely correlated with basal ROS levels in EGFR mutation–positive NSCLC cell lines. Knockdown of CD44v induced depletion of intracellular GSH and increased ROS levels in EGFR-mutated NSCLC cells that express CD44v at a high level (CD44vhigh). In addition, depletion of GSH by treatment with buthionine-[S, R]-sulfoximine induced marked accumulation of ROS and enhanced the cytotoxicity of cisplatin in CD44vhigh EGFR-mutated NSCLC cells but not in corresponding CD44vlow cells. This enhancement of cisplatin cytotoxicity by GSH depletion was prevented by treatment with the antioxidant N-acetyl-L-cysteine. Knockdown of CD44v also enhanced cisplatin cytotoxicity in CD44vhigh EGFR mutation–positive NSCLC cells but not in CD44vlow cells. Conclusion Our results thus implicate CD44v in redox adaptation and as a potential target for treatment in CD44vhigh EGFR-mutated NSCLC cells.

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