Inhibitory Effect of Epidermal Growth Factor on the Proliferation of Lung Cancer Cell Lines

SeungHee Lee; Hong-Gyun Wu

doi:10.6058/jlc.2010.9.2.64

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Lee and Wu: Inhibitory Effect of Epidermal Growth Factor on the Proliferation of Lung Cancer Cell Lines

Original Article

J Lung Cancer 2010;9(2):64-71.

Published online: 10 January 2010

DOI: https://doi.org/10.6058/jlc.2010.9.2.64

Inhibitory Effect of Epidermal Growth Factor on the Proliferation of Lung Cancer Cell Lines

SeungHee Lee, M.S., Hong-Gyun Wu, M.D., Ph.D.

Department of Radiation Oncology & Cancer Research Institute, Seoul National University College of Medicine and Institute of Radiation Medicine, Medical Research Center, Seoul National University, Seoul, Korea

Address for correspondence Hong-Gyun Wu, M.D., Ph.D. Department of Radiation Oncology, Seoul National University College of Medicine, 28, Yeongeon-dong, Jongno-gu, Seoul 110-744, Korea Tel: 82-2-2072-3177 Fax: 82-2-765-3317 E-mail: wuhg@snu.ac.kr

This work supported by research grant from Korean Association for the Study of Lung Cancer in 2008 and grant No. 2009–0083886 from the Korea Science and Engineering Foundation grant funded by the Korea government (MEST).

Received 29 July 2010 Revised 9 August 2010 Accepted 19 August 2010

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Purpose

Epidermal growth factor (EGF) is known to cause cellular proliferation, differentiation and survival. However, there are a few articles that have reported on the cell killing effect of EGF. We evaluated the effects of EGF on the survival of some lung cancer cell lines and we tried to determine the mechanism of action.

Materials and Methods

We examined various lung cancer cell lines. The cultured cells were exposed to radiation alone (0, 2, 5, and 10 Gy), EGF alone (0∼1,000 nM) or a combination of radiation and EGF (10 nM). Survival was measured using a clonogenic assay and the expressions of epidermal growth factor receptor (EGFR), Ki-67 and cleaved-PARP were detected by western blot analysis. K-ras mutations were detected using polymerase chain reaction and sequencing.

Results

Treatment of EGF induced cell death in the lung cancer cell lines. The addition of EGF (10 nM) to radiation (0, 2, 5, and 10 Gy) resulted in an increased cell killing effect of radiation. The EGFR expression decreased with the addition of EGF. EGF increased the expression of cleaved-PARP, but it did not increase the expression of Ki-67. The effects of EGF were not correlated with EGFR mutation or K-ras mutation.

Conclusion

In our study, EGF inhibited the proliferation of lung cancer cell lines and it induced apoptosis. EGF enhanced the radiosensitivity of lung cancer cells when EGF was combined with radiation. It is suggested that EGF seem to be one of the cytotoxic agents for lung cancer cell lines.

Keywords: Epidermal growth factor, Radiation, Apoptosis, Lung cancer cell lines

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Figures

Fig. 1.

The effect of epidermal growth factor (EGF) on the proliferation of the lung cancer cell lines. The cells were cultured for 24 hours prior to treatment with various concenrations (0, 0.01, 0.1, 1, 10, 100, and 1,000 nM) of EGF. The surviving cells were counted on day 10 using a clonogenic assay.

Fig. 2.

The effect of epidermal growth factor (EGF) on the proliferation of lung cancer cell lines. The cells were growth-arrested by incubation in medium containing 0.05% serum for 1 day. The cells were cultured for 12 days with various concenrations (0, 0.01, 0.1, 1, 10, 100, and 1,000 nM) of EGF.

Fig. 3.

Clonogenic survival of the lung cancer cells treated with epidermal growth factor (EGF) followed by irradiation with a range of radiation doses. Although the radiation sensitizing effect was minimal in the A549 cell line, the rest of lung cancer cell lines showed the radiation sensitizing effect of EGF.

Fig. 4.

Expression of epidermal growth factor receptor (EGFR) in various cancer cell lines.

Fig. 5.

The epidermal growth factor receptor (EGFR), Ki-67 and cleaved PARP expressions in response to epidermal growth factor (EGF) treatment on the lung cancer cell lines. Treatment with EGF decreased the expression of EGFR and it increased the expression of cleaved PARP.

Fig. 6.

Mutation analysis of K-ras in the lung cancer cell lines.

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