In this pro-oxidant scenario, the ADP produced by high HK activity could control both membrane potential and ROS Afatinib generation through an ADP-recycling NSC 136476 mechanism similar to that proposed by Da-Silva et al. In connection with the mechanism of NaB activation of respiration, various interpretations, not necessarily mutually excluding, are possible. In one scenario, there could be a direct interaction of the HDACi with any of the mitochondrial components, similarly to what was described for HK. Conversely, NaB could act elsewhere and ultimately affect the respiratory parameters of the mitochondria. Perhaps the broadest consideration that must be taken into account is the possibility that most if not all the HDACis effects described in the present work could have been brought about by an action on a central factor that, in turn, would affect other pathways. One possibility is that HDACis are decreasing HIF-1a, and this effect decreases glycolysis maintained by HIF-1a. A key transcription factor that knowingly acts as the hub for many processes associated to the cell cycle and the energy metabolism itself. Thus, by preferentially targeting HIF-1a, NaB or TSA could indirectly affect hundreds of other genes. At any rate, investigation of the effect of HDACis on HIF1a regulation and on key actors of anaerobic and oxidative metabolism is currently under way. Although more evidence has yet to be procured, the set of results contained herein suggests that NaB and TSA could induce H460 cells to assume an altogether differentiated state. It is proposed that in this differentiated phenotype, cells actually shift towards more oxidative metabolism akin to untransformed cells. Ultimately, the enhanced oxidative metabolism would harm the H460 cells by way of excessive production of ROS that can cause cell death. In this context, the H460 cells treated with NaB could activate antioxidant defense mechanisms related to the increase in: HK activity bound to mitochondria, which increases ADP recycling; G6PDH activity, which increases PPP flux and NADPH supply; and the expression of Mfn 1, which, may reflect the initial stages of a process of mitochondrial fusion that can ultimately act as inhibitor of apoptosis. Indeed it has been shown that under oxidative stress mitochondria can rescue their membrane potential. Although the notion that NaB and TSA induced generation of ROS is a reasonable proposition, it must be mentioned that the topic of ROS and cancer cells is in itself quite controversial. There are data supporting the view that ROS are in fact essential requirements for the survival of tumor cells, even though the mechanism whereby ROS would support tumor growth remains elusive. Taken together the results presented here revealed a unique biochemical profile induced by NaB and TSA. The data essentially showed that alterations in the glycolytic flux triggered an enhancement of mitochondrial function that was not, however, paralleled by cell proliferation. Whilst further studies are paramount to unveil the mechanisms underlying the interplay between glycolysis and oxidative metabolism, the groundwork established in the present work already suggests that the stimulation of the oxidative metabolism in tumor cells may be an interesting strategy for chemotherapy. Ligand-based virtual screening, quantitative structureproperty and structure-activity relationships, and other concepts in computational medicinal chemistry are based on the similarity principle, which states that similar compounds generally exhibit similar properties.