4EASO transfection suppresses mesothelioma proliferation.

<p>Normal mesothelial cells (LP9) and mesothelioma cell lines H2373, H2461 and H2596 were treated with the indicated concentration of mmASO (open columns) and 4EASO (filled columns), and viable cells were counted. Normal LP9 cells were most resistant to 4EASO treatment, while growth of mesothelioma cell lines was reduced extensively. <i>Columns</i>, the mean of three independent determinations of cell number normalized to untreated cells, <i>bars</i>, s.d.</p

4EASO transfection induces apoptosis in mesothelioma.

<p>Mesothelioma cell lines H2373 and H2461 along with human mesothelial cells (LP9) were treated with the indicated concentration of 4EASO for 48 hours. Lysates were immunoblotted with anti-PARP antibody. In mesothelioma cell lines PARP cleavage was substantially increased in 4EASO treated compared to untreated cells.</p

eIF4E expression is reduced by 4EASO treatment in mesothelioma.

<p>Cultured normal mesothelial and mesothelioma cells were transfected with mmASO or 4EASO and eIF4E and β-actin protein expression was evaluated by immunoblot analysis from lysates harvested after 72 hours. The mmASO does not alter eIF4E levels in mesothelial cells or mesothelioma cells. The band intensity levels for eIF4E following 4EASO treatment was normalized to untreated cells for each cell line and was determined using ImageJ. β-actin level does not change upon treatment and serves as a loading control.</p

Enhanced susceptibility of mesothelioma cells treated with 4EASO to cytotoxic drugs.

<p>Mesothelioma cell lines transfected with mmASO or 4EASO were treated with the indicated concentration of gemcitabine (GEM) or pemetrexed (PEM) and viable cells were counted. Elevated cell death was found for cells treated in combination with 4EASO and pemetrexed or gemcitabine compared to each treatment alone. Concurrent treatment of mesothelioma cells with 4EASO combined with pemetrexed or gemcitabine suppresses proliferation compared to treatment with mmASO combined with pemetrexed or gemcitabine. <i>Columns</i>, the mean of three independent determinations of cell number normalized to untreated cells, <i>bars</i>, s.d. Averages of combination treatment was compared to either agent alone by Student’s <i>t</i>-test. * denotes a p value <0.05. NS = not statistically significant.</p