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 language | English |
|---|---|
| Pages (from-to) | 488-493 |
| Number of pages | 6 |
| Journal | Clinical Lung Cancer |
| Volume | 13 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - Nov 2012 |
Fingerprint
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 journal › Article
}
TY - JOUR
T1 - Knockdown of the epidermal growth factor receptor gene to investigate its therapeutic potential for the treatment of non-small-cell lung cancers
AU - Shien, Kazuhiko
AU - Ueno, Tsuyoshi
AU - Tsukuda, Kazunori
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
PY - 2012/11
Y1 - 2012/11
N2 - 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.
AB - 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.
KW - Epidermal growth factor receptor
KW - Epidermal growth factor receptor-tyrosine kinase inhibitors
KW - Non-small-cell lung cancer
KW - Oncogene addiction
KW - Small interfering RNA
UR - http://www.scopus.com/inward/record.url?scp=84867604691&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84867604691&partnerID=8YFLogxK
U2 - 10.1016/j.cllc.2012.02.003
DO - 10.1016/j.cllc.2012.02.003
M3 - Article
C2 - 22503506
AN - SCOPUS:84867604691
VL - 13
SP - 488
EP - 493
JO - Clinical Lung Cancer
JF - Clinical Lung Cancer
SN - 1525-7304
IS - 6
ER -