Enhancement of Apo2L/TRAIL (tumor necrosis factor–related apoptosis-inducing ligand)–induced apoptosis in non–small cell lung cancer cell lines by chemotherapeutic.

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Enhancement of Apo2L/TRAIL (tumor necrosis factor–related apoptosis-inducing ligand)–induced apoptosis in non–small cell lung cancer cell lines by chemotherapeutic agents without correlation to the expression level of cellular protease caspase-8 inhibitory protein Steffen Frese, MDa, Thomas Brunner, PhDb, Mathias Gugger, MDb, Aima Uduehi, PhDa, Ralph A. Schmid, MDa The Journal of Thoracic and Cardiovascular Surgery Volume 123, Issue 1, Pages 168-174 (January 2002) DOI: 10.1067/mtc.2002.119694 Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 1 Apo2L/TRAIL induces apoptosis in NSCLC cell lines, but this is not correlated with expression of cFLIP. A, Cells were treated with 1-mg/mL TRAIL/Apo2L for 24 hours, and apoptosis was assessed by annexin V/propidium iodide staining followed by FACScan analysis (n = 3). Bars represent mean; error bars represent SD. B, Expression of c-FLIP in NSCLC cell lines was determined by Western blot analysis. To confirm equal protein levels the same blot was stripped and developed with antitubulin antibody. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 168-174DOI: (10.1067/mtc.2002.119694) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 2 Apo2L/TRAIL-induced apoptosis is augmented by chemotherapeutic agents. Cells were incubated for 24 hours with 100ng/mL Apo2L/TRAIL (white bars), with chemotherapeutic agents (gray bars), and with combined Apo2L/TRAIL and chemotherapy (black bars). Apoptosis was measured by annexin V/propidium iodide staining followed by flow cytometry (n = 3). Bars represent mean; error bars represent SD. Concentrations of chemotherapeutic agents are indicated with numbers: 1 and 2, 0.5-mmol/L and 5-mmol/L doxorubicin; 3 and 4, 3-mmol/L and 30-mmol/L 5-fluorouracil; 5 and 6, 0.18-mmol/L and 1.8-mmol/L camptothecin; 7 and 8, 10-μmol/L and 100-μ mol/L cisplatin; 9 and 10, 10-nmol/L and 100-nmol/L paclitaxel. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 168-174DOI: (10.1067/mtc.2002.119694) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 3 Augmentation of Apo2L/TRAIL-induced apoptosis does not correlate with expression of c-FLIP. Cells were incubated with 5-mmol/L doxorubicin, 30-mmol/L 5-fluorouracil, 1.8-mmol/L camptothecin, 100-μmol/L cisplatin, 100-nmol/L paclitaxel, and 1-μg/mL cycloheximide for 24 hours. Expression of c-FLIP was determined by Western blot. Equal amount of protein was shown by reblotting against tubulin. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 168-174DOI: (10.1067/mtc.2002.119694) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions

Fig. 4 Expression of cFLIP in primary NSCLC (C) and normal lung tissue (N). Protein lysates were obtained from frozen tissue from patients who underwent lung resection. c-FLIP expression was determined by Western blot. To confirm equal protein levels the same blot was stripped and developed with antitubulin antibody. A, Adenocarcinoma; B, squamous cell carcinoma. The Journal of Thoracic and Cardiovascular Surgery 2002 123, 168-174DOI: (10.1067/mtc.2002.119694) Copyright © 2002 American Association for Thoracic Surgery Terms and Conditions