Combination Therapy of Bortezomib (PS-341) and SAHA (Vorinostat) in Non-Small Cell Lung Cancer Cell Lines

Abstract

Purpose:The recent introduction of targeted therapy for non-small cell lung cancer (NSCLC) has changed the paradigm of lung cancer chemotherapy. However, only a small portion of NSCLC patients received the benefit of these new drugs. A proteasome inhibitor (bortezomib) and a histone deacetylase inhibitor (SAHA) were approved for clinical use for treating some hematologic malignancies. In this study, we investigated the combination treatment of bortezomib and SAHA in NSCLC cell lines. Materials and Methods: The combined effects of bortezomib and SAHA on lung cancer cell lines were measured by Calcusyn software. Induction of apoptosis was examined by performing an Annexin V assay. Generation of reactive oxygen species (ROS) and protection by N-acetylcysteine were measured by flow cytometry after staining with DCFH-DA. The effect of the combination of drugs on apoptosis and autophagy was investigated by Western blot assay. Results: Strong synergism was found for the combination of bortezomib and SAHA. The synergistic interaction was mediated by strong apoptosis. Increased ROS generation was partly responsible for the induction of apoptosis, and this was suppressed by the ROS scavenger N-acetylcysteine. Combined treatment induced the strong activation of caspase-3 and the breakdown of the antiapoptotic molecule Bcl-2. Furthermore, increased breakdown of beclin-1, which is known to be an autophagic molecule, was also found. Conclusion: Combination therapy with bortezomib and SAHA showed a strong synergistic antitumor effect on human lung cancer cell lines. Enhanced induction of apoptosis was a responsible mechanism, and this was partly mediated by ROS generation. Further studies are warranted for determining the role of apoptosis and autophagy for this combination therapy. Medicine, 300,