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Pemetrexed downregulates ERCC1 expression and enhances cytotoxicity effected by resveratrol in human nonsmall cell lung cancer cells

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Abstract

The multitargeted antifolate pemetrexed has demonstrated certain clinical activities against nonsmall cell lung cancer (NSCLC). Resveratrol (3,5,4-trihydroxy-trans-stilbene) is a polyphenol found in grapes and other plants and has great potential as a preventative and therapeutic agent due to its anticarcinogenic activity. The efficacy of adding resveratrol to pemetrexed to prolong the survival of patients with NSCLC still remains unclear. The excision repair cross-complementation 1 (ERCC1) is a DNA repair gene coding 5′ endonuclease in nucleotide excision repair and is overexpressed in chemo- or radioresistant carcinomas. In this study, resveratrol (10–50 μM) inhibited cell survival in two NSCLC cells, H520 and H1975. Treatment with resveratrol increased ERCC1 messenger RNA and protein levels in a MKK3/6-p38 MAPK signal activation-dependent manner. Furthermore, blocking p38 MAPK activation by SB202190 or knocking down ERCC1 expression by transfection with small interfering RNA of ERCC1 enhanced the cytotoxicity of resveratrol. Combining resveratrol with pemetrexed resulted in a synergistic cytotoxic effect, accompanied with the reduction of phospho-p38 MAPK and ERCC1 protein levels, and a DNA repair capacity. Expression of constitutively active MKK6 (MKK6E) or HA-p38 MAPK vectors significantly rescued the decreased p38 MAPK activity, and restored ERCC1 protein levels and cell survival in resveratrol and pemetrexed cotreated NSCLC cells. In this study, for the first time, we have demonstrated the synergistic effect of combined treatment with resveratrol and pemetrexed in human NSCLC cells through downregulation of the MKK3/6-p38 MAPK-ERCC1 signal, suggesting a potential benefit of combining resveratrol and pemetrexed to treat lung cancer in the future.

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Abbreviations

CFA:

Colony-forming ability

ERCC1:

Excision repair cross-complementation 1

ERK:

Extracellular signal-regulated kinase

siRNA:

Small interfering RNA

MAPK:

Mitogen-activated protein kinase

NSCLC:

Nonsmall cell lung cancer

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Acknowledgments

We thank Dr. Tsui-Chun Tsou, Dr. Show-Mei Chuang, and Dr. Jia-Ling Yang for providing us with expression plasmids for transfection. Grant supports from the National Science Council, Taiwan, grant NSC 99-2320-B-415-001-MY3 and NSC 102-2320-B-415-006 (Y-W. Lin), and National Taiwan University Hospital, Hsin-Chu Branch, Taiwan (grant HCH101-05, HCH102-02) are acknowledged.

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The authors declare that they have no conflict of interest.

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Correspondence to Yun-Wei Lin.

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R.-S. Chen, J.-C. Ko, and H.-C. Chiu contributed equally.

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Chen, RS., Ko, JC., Chiu, HC. et al. Pemetrexed downregulates ERCC1 expression and enhances cytotoxicity effected by resveratrol in human nonsmall cell lung cancer cells. Naunyn-Schmiedeberg's Arch Pharmacol 386, 1047–1059 (2013). https://doi.org/10.1007/s00210-013-0905-9

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