Indirubin-3-monoxime and thymoquinone exhibit synergistic efficacy as therapeutic combination in in-vitro and in-vivo models of Lung cancer

In this study, we report the combination of indirubin-3-monoxime (I3M) and thymoquinone (Tq) to have excellent therapeutic efficacy in models of Lung cancer (LC). Preliminary screening was done with A549 cells. Cell cycle, apoptosis and NF kappa B phosphorylation were determined by flow cytometry, while apoptotic proteins, Akt and mTOR were assessed by western blotting. Mouse xenograft model was used to assess the therapeutic efficacyin-vivo. Synergistic reduction in cell viability was observed with I3M + Tq combinations, which were non-toxic to normal HFL-1 cells. Cell cycle analysis indicated G(1)phase reduction with subsequent accumulation of sub G(0)contents. Annexin V assay revealed higher apoptotic cells with combinations compared to individual treatments with a decrease in Bcl-2/Bax ratio. The combinations exhibited anti-metastasis activity in cell migration in the scratch, scatter and tumour cell migration assays and effectively reduced the tumour growth in mouse xenograft model. Expression levels of p-AKT, p-mTOR, Caspase-3, p-53 and NF kappa B were significantly reduced in the combination treated mice compared to individual treatments. Results of current study demonstrate clear efficacy of I3M + Tq combinations in LC models mediated by suppressing Akt/mTOR/NF kappa B signalling. Further research is recommended to transform these findings into novel therapeutic combinations against LC.