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 NFκB 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 efficacy in-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 phase reduction with subsequent accumulation of sub G 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 NFκB 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/NFκB signalling. Further research is recommended to transform these findings into novel therapeutic combinations against LC.