Synergism of a novel MCL-1 downregulator, acriflavine, with navitoclax (ABT-263) in triple-negative breast cancer, lung adenocarcinoma and glioblastoma multiforme

Myeloid cell leukemia sequence 1 (MCL-1), an anti-apoptotic B-cell lymphoma 2 (BCL-2) family molecule frequently amplified in various human cancer cells, is known to be critical for cancer cell survival. MCL-1 has been recognized as a target molecule for cancer treatment. While various agents have emerged as potential MCL-1 blockers, the present study presented acriflavine (ACF) as a novel MCL-1 inhibitor in triple-negative breast cancer (TNBC). Further evaluation of its treatment potential on lung adenocarcinoma and glioblastoma multiforme (GBM) was also investigated. The anticancer effect of ACF on TNBC cells was demonstrated when MDA-MB-231 and HS578T cells were treated with ACF. ACF significantly induced typical intrinsic apoptosis in TNBCs in a dose- and time-dependent manner via MCL-1 downregulation. MCL-1 downregulation by ACF treatment was revealed at each phase of protein expression. Initially, transcriptional regulation via reverse transcription-quantitative PCR was validated. Then, post-translational regulation was explained by utilizing an inhibitor against protein biosynthesis and proteasome. Lastly, immunoprecipitation of ubiquitinated MCL-1 confirmed the post-translational downregulation of MCL-1. In addition, the synergistic treatment efficacy of ACF with the well-known MCL-1 inhibitor ABT-263 against the TNBC cells was explored [combination index (CI)<1]. Conjointly, the anticancer effect of ACF was assessed in GBM (U87, U251 and U343), and lung cancer (A549 and NCI-H69) cell lines as well, using immunoblotting, cytotoxicity assay and FACS. The effect of the combination treatment using ACF and ABT-263 was estimated in GBM (U87, U343 and U251), and non-small cell lung cancer (A549) cells likewise. The present study suggested a novel MCL-1 inhibitory function of ACF and the synergistic antitumor effect with ABT-263.