Anti-cancer effect of celecoxib and aerosolized docetaxel against human non-small cell lung cancer cell line, A549

Abstract

Direct delivery of chemotherapeutic agents to the lung can increase both the drug concentration and exposure period to lung tumours. The objective of this study was to formulate docetaxel (DOC) into a metered dose inhaler (MDI), assess its aerodynamic characteristics and to evaluate the effect of celecoxib (CXB), a cyclooxygenase-2 (COX-2) inhibitor, on the in-vitro cytotoxicity and apoptotic response of aerosolized DOC against human lung adenocarcinoma cell line A549. A stable solution-type MDI formulation was developed with 0.25% DOC and 15% w/w ethyl alcohol using HFA 134a propellant. The formulation was evaluated for medication delivery, mass median aerodynamic diameter (MMAD), geometric standard deviation (GSD), percent throat deposition, respirable mass and respirable fraction. A six-stage viable impactor was used to assess the in-vitro cytotoxicity of DOC-MDI alone or in combination with CXB. Induction of apoptosis in A549 cells by DOC (non-aerosolized and aerosolized) in combination with CXB was evaluated by established techniques, such as caspase-3 estimation and terminal deoxynucleotidyl transferase-mediated nick end labelling (TUNEL) staining. The influence of different treatments on the expression of COX-2 and peroxisome proliferator-activated receptor-γ (PPAR-γ) in A549 cells was studied by RT-PCR. The DOC-MDI formulation had a MMAD of 1.58 μm, (GSD = 3.2) and a medication delivery of 80 μg/shot. DOC-MDI (one shot) in combination with CXB (10 μg mL−1) had a cell kill of more than 80% as determined by in-vitro cytotoxicity assay. The specific caspase-3 activity in A549 cells treated with DOC (0.01 μg mL−1) and CXB (10.0 μg mL−1) combination was 4 times higher than CXB and untreated control group, respectively. Further, TUNEL staining showed significant apoptosis of A549 cells treated with aerosolized DOC alone or in combination with CXB when compared with CXB and untreated cells. The RT-PCR experiments showed similar expression of COX-2 in both control and treated groups. PPAR-γ expression was increased in the combination treatment (0.01 μg mL−1 DOC and 10 μg mL−1 CXB) as compared with control (untreated), DOC (0.01 μg mL−1) and CXB (10 μg mL−1) treatments. Our results indicate the potential of inhalation delivery of DOC in the treatment of lung cancer.