Oncotarget

Research Papers:

The impact of intermittent versus continuous exposure to EGFR tyrosine kinase inhibitor on selection of EGFR T790M-mutant drug-resistant clones in a lung cancer cell line carrying activating EGFR mutation

Youngjoo Lee _, Yu-Ra Choi, Kyoung-Yeon Kim and Dong Hoon Shin

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Oncotarget. 2016; 7:43315-43323. https://doi.org/10.18632/oncotarget.9703

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Abstract

Youngjoo Lee1,2, Yu-Ra Choi2, Kyoung-Yeon Kim2, Dong Hoon Shin2

1Center for Lung Cancer, National Cancer Center, Goyang, Republic of Korea

2Lung Cancer Branch, Research Institute and Hospital, National Cancer Center, Goyang, Republic of Korea

Correspondence to:

Youngjoo Lee, email: yjlee@ncc.re.kr

Keywords: drug resistance, drug sensitivity, EGFR tyrosine kinase inhibitor, EGFR mutation, non-small-cell lung cancer

Received: November 09, 2015     Accepted: May 09, 2016     Published: May 30, 2016

ABSTRACT

Drug-resistant cell lines are essential tools for investigating the mechanisms of resistance to molecular-targeted anti-cancer drugs. However, little is known about how to establish clinically relevant drug-resistant cell lines. Our study examined the impact of a drug-free period on the establishment of a cell line with clinically relevant resistance to molecular-targeted drugs. We used PC9 cells, a lung cancer cell line carrying EGFR mutation, because this is a validated target for EGFR tyrosine kinase inhibitors (TKI). PC9 cells were intermittently or continuously exposed to increasing concentrations of gefitinib (0.01 μM to 1.0 μM) and the emergence of the most common acquired resistance mutation in EGFR, T790M, was determined. T790M was detected at a 25-fold lower drug concentration in cells continuously exposed to gefitinib (PC9/GRc) than in cells intermittently exposed to gefitinib (PC9/GRi) (0.04 μM vs 1.0 μM, respectively). The mutation frequencies at those drug concentrations were 19.8% and 8.0% in PC9/GRc and PC9/GRi cells, respectively. After drug-free culture for 8 weeks, resistance to gefitinib decreased in the PC9/GRi cells but not in the PC9/GRc cells. In the PC9/GRc cells, the frequency of the T790M mutation was consistently about 20% from 0.04 μM to 1.0 μM of gefitinib. In the PC9/GRc cells, the T790M mutation was detected in all single-cell clones, at frequencies ranging from 7.0% to 37.0%, with a median of 19.5% (95% confidence interval, 17.3%–20.9%). In conclusion, compared with intermittent drug exposure, continuous exposure might select better minor drug-resistant clones when creating cell lines resistant to molecular-targeted drugs.


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