Knockdown of the epidermal growth factor receptor gene to investigate its therapeutic potential for the treatment of non-small-cell lung cancers

Kazuhiko Shien, Tsuyoshi Ueno, Kazunori Tsukuda, Junichi Sou, Kenichi Suda, Takafumi Kubo, Masashi Furukawa, Takayuki Muraoka, Yuho Maki, Norimitsu Tanaka, Hiromasa Yamamoto, Katsuyuki Kiura, Tetsuya Mitsudomi, Shinichi Toyooka, Shinichiro Miyoshi

Research output: Contribution to journalArticle

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Abstract

Background: Epidermal growth factor receptor (EGFR) is often overexpressed in non-small-cell lung cancer (NSCLC). Anti-EGFR agents, including EGFR-tyrosine kinase inhibitors are considered to be effective when a drug-sensitive EGFR mutation is present. However, inherent and acquired resistances are major problems of EGFR-targeting therapies. In this study, we performed EGFR knockdown by using small interfering RNAs in NSCLC cell lines to examine the significance of targeting EGFR for NSCLC therapy. Methods: We treated 13 NSCLC cell lines, including 8 EGFR mutant and 5 EGFR wild type by using gefitinib or small interfering RNAs against EGFR (siEGFR). Three cell lines (PC-9-GR1, RPC-9, and HCC827-ER) were experimentally established with acquired resistance to EGFR-tyrosine kinase inhibitors. The antitumor effect was determined by using an 3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H- tetrazolium, inner salt (MTS) or colony formation assay. The protein expression was evaluated by using Western blotting. Results: All 13 cell lines expressed EGFR protein, and siEGFR downregulated EGFR protein expression in all. The cell viability was suppressed by siEGFR in 6 of 8 EGFR-mutant cell lines (suppressed 57%-92% of control cells), including PC-9-GR1 and RPC-9. The NCI-H1650 and HCC827-ER harbored EGFR mutations but were not suppressed. Of note, PTEN (phosphatase and tensin homolog) was deleted in NCI-H1650, and c-MET was amplified in HCC827-ER. It was not suppressed in any of the EGFR wild-type cells except in the NCI-H411, in which EGFR is phosphorylated, which indicates its activation. Conclusions: Analysis of the results indicated that EGFR can be a therapeutic target in NSCLCs with EGFR activation. In contrast, targeting EGFR is not appropriate for tumors in which EGFR is not activated, even if EGFR is expressed.

Original languageEnglish
Pages (from-to)488-493
Number of pages6
JournalClinical Lung Cancer
Volume13
Issue number6
DOIs
Publication statusPublished - Nov 2012

Fingerprint

erbB-1 Genes
Epidermal Growth Factor Receptor
Non-Small Cell Lung Carcinoma
Therapeutics
Small Interfering RNA
Cell Line
Protein-Tyrosine Kinases

Keywords

  • Epidermal growth factor receptor
  • Epidermal growth factor receptor-tyrosine kinase inhibitors
  • Non-small-cell lung cancer
  • Oncogene addiction
  • Small interfering RNA

ASJC Scopus subject areas

  • Cancer Research
  • Oncology
  • Pulmonary and Respiratory Medicine

Cite this

Knockdown of the epidermal growth factor receptor gene to investigate its therapeutic potential for the treatment of non-small-cell lung cancers. / Shien, Kazuhiko; Ueno, Tsuyoshi; Tsukuda, Kazunori; Sou, Junichi; Suda, Kenichi; Kubo, Takafumi; Furukawa, Masashi; Muraoka, Takayuki; Maki, Yuho; Tanaka, Norimitsu; Yamamoto, Hiromasa; Kiura, Katsuyuki; Mitsudomi, Tetsuya; Toyooka, Shinichi; Miyoshi, Shinichiro.

In: Clinical Lung Cancer, Vol. 13, No. 6, 11.2012, p. 488-493.

Research output: Contribution to journalArticle

Shien, K, Ueno, T, Tsukuda, K, Sou, J, Suda, K, Kubo, T, Furukawa, M, Muraoka, T, Maki, Y, Tanaka, N, Yamamoto, H, Kiura, K, Mitsudomi, T, Toyooka, S & Miyoshi, S 2012, 'Knockdown of the epidermal growth factor receptor gene to investigate its therapeutic potential for the treatment of non-small-cell lung cancers', Clinical Lung Cancer, vol. 13, no. 6, pp. 488-493. https://doi.org/10.1016/j.cllc.2012.02.003
Shien, Kazuhiko ; Ueno, Tsuyoshi ; Tsukuda, Kazunori ; Sou, Junichi ; Suda, Kenichi ; Kubo, Takafumi ; Furukawa, Masashi ; Muraoka, Takayuki ; Maki, Yuho ; Tanaka, Norimitsu ; Yamamoto, Hiromasa ; Kiura, Katsuyuki ; Mitsudomi, Tetsuya ; Toyooka, Shinichi ; Miyoshi, Shinichiro. / Knockdown of the epidermal growth factor receptor gene to investigate its therapeutic potential for the treatment of non-small-cell lung cancers. In: Clinical Lung Cancer. 2012 ; Vol. 13, No. 6. pp. 488-493.
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AU - Sou, Junichi

AU - Suda, Kenichi

AU - Kubo, Takafumi

AU - Furukawa, Masashi

AU - Muraoka, Takayuki

AU - Maki, Yuho

AU - Tanaka, Norimitsu

AU - Yamamoto, Hiromasa

AU - Kiura, Katsuyuki

AU - Mitsudomi, Tetsuya

AU - Toyooka, Shinichi

AU - Miyoshi, Shinichiro

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