Formulation and Evaluation of Aerosolized Celecoxib for the Treatment of Lung Cancer
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- Haynes, A., Shaik, M., Chatterjee, A. et al. Pharm Res (2005) 22: 427. doi:10.1007/s11095-004-1881-z
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We examined the effect of aerosolized celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, on the in vitro cytotoxicity and apoptotic response of docetaxel against the human lung carcinoma cell lines A549 and H460.
A metered dose inhaler (MDI) formulation of celecoxib was prepared and evaluated for its medication delivery and aerodynamic properties. The in vitro cytotoxicity of the aerosolized celecoxib-MDI alone or in combination with docetaxel was assessed using a six-stage viable impactor by a previously established method. The induction of apoptosis was evaluated by morphologic examination (acridine orange and Hoechst staining) and DNA fragmentation. Furthermore, in an attempt to identify molecular targets involved in the anticancer mechanisms of celecoxib and docetaxel, we examined their effect on the expression of an array of markers involved in the COX-2 dependent a nd independent pathways.
The celecoxib-MDI had a medication delivery of 231.3 μg/shot, mass median aerodynamic diameter (MMAD) of 1.4 μm (GSD=1.9), and respirable fraction of 50.7%. The celecoxib-MDI (2 shots) in combination with docetaxel had cell kills as high as 81.3% and 67.7% in A549 and H460 cells, respectively. Hoechst and acridine orange staining showed an enhanced induction of apoptosis in A549 and H460 cells exposed to aerosolized celecoxib with docetaxel, which was further confirmed by DNA fragmentation. Western blot analysis showed a significant reduction in cPLA2 expression in both A549 and H460 cells treated with the combination of celecoxib with docetaxel. In the COX-2 independent pathway, there was a significant increase in the expression of PPAR-γ and p53, whereas pro-caspase-3 expression was significantly decreased, which may contribute to the enhanced apoptotic response observed with the combination treatment.
Our results suggest that aerosolized celecoxib significantly enhances the in vitro cytotoxicity and apoptotic response of docetaxel against A549 and H460 cells, and this enhanced activity is mediated via alterations in expression of various molecular targets involved in apoptosis.
Key Words:napoptosis celecoxib cytotoxicity docetaxel inhalation
cytosolic phospholipase A2
geometric standard deviation
hydrofluoroalkane; 5-LOX, 5-lipoxygenase
medication delivery device
metered dose inhaler
mass median aerodynamic diameter
nonsteroidal anti-inflammatory drug
non-small cell lung cancer
peroxisome proliferator-activated receptor
tumor suppressor protein