Histone Deacetylase Inhibitor Romidepsin Enhances Anti-Tumor Effect of Erlotinib in Non-small Cell Lung Cancer (NSCLC) Cell Lines Wei Zhang, PhD, Michael.

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Histone Deacetylase Inhibitor Romidepsin Enhances Anti-Tumor Effect of Erlotinib in Non-small Cell Lung Cancer (NSCLC) Cell Lines Wei Zhang, PhD, Michael Peyton, PhD, Yang Xie, PhD, Junichi Soh, MD, John D. Minna, MD, Adi F. Gazdar, MD, Eugene P. Frenkel, MD Journal of Thoracic Oncology Volume 4, Issue 2, Pages 161-166 (February 2009) DOI: 10.1097/JTO.0b013e318194fae7 Copyright © 2009 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 1 Romidepsin synergistically increased the sensitivity of erlotinib in non-small cell lung cancer (NSCLC) cell lines. A, Nine NSCLC cell lines were treated with varying concentrations of erlotinib either in the absence or presence of romidepsin (1 ng/ml) for 72 hours. Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and representative pairs of cell viability curves are shown. B, The dose-normalized isobologram for erlotinib and romidepsin with normalization of dose with IC50 to unity on both x and y axis. All the points represent the drug combination that yield 50% cell reduction. If the point falls on the lower left part of the graph, synergism is indicated. Journal of Thoracic Oncology 2009 4, 161-166DOI: (10.1097/JTO.0b013e318194fae7) Copyright © 2009 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 2 Erlotinib and romidepsin induced apoptosis in non-small cell lung cancer (NSCLC) cell lines. NCI-H1299 cell line (A) and NCI-H23 cell line (B) treated with 5 μM erlotinib or/and 1 ng/ml romidepsin were analyzed for apoptosis using a cell death detection kit (Roche). Results represent as means ± SD of three independent experiments. *p < 0.05 versus control group (Student's t test). Journal of Thoracic Oncology 2009 4, 161-166DOI: (10.1097/JTO.0b013e318194fae7) Copyright © 2009 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 3 Growth inhibition of NCI-H1299 cell line xenografts by erlotinib and romidepsin coadministration. Five × 106 NCI-H1299 cells were injected subcutaneously into each of twenty female BALB/c athymic nude mice. These mice were divided into four groups at day 7 after tumor development. They were injected with either 1 × phosphate-buffered saline (PBS), romidepsin alone, erlotinib alone or the combination. Romidepsin was administered 3 times at 4-day intervals (1.2 mg/kg body weight). Erlotinib was administered 5 days a week (50 mg/kg body weight). Tumor sizes were measured at the indicated days. Results are shown as means + SEM of groups of 5 mice. Statistical significance was determined by Student's t test (*p <0.05 versus control). Journal of Thoracic Oncology 2009 4, 161-166DOI: (10.1097/JTO.0b013e318194fae7) Copyright © 2009 International Association for the Study of Lung Cancer Terms and Conditions