Abstract
SummarySignal transduction pathways, which regulate cell growth and survival, are up-regulated in many cancers and there is considerable interest in their pharmaceutical modulation for cancer treatment. However inhibitors of single pathway components induce feedback mechanisms that overcome the growth moderating effect of the inhibitor. Combination treatments have been proposed to provide a more complete pathway inhibition. Here the effect of dual treatment of cancer cells with a pan-Akt and a pan-mTOR inhibitor was explored. Breast (SKBr3 and MDA-MB-468) and colorectal (HCT8) cancer cells were treated with the pan-Akt inhibitor MK2206 and pan-mTOR inhibitor AZD8055. Cytotoxic effect of the two drugs were determined using the MTT assay and the Combination Index and isobolomic analysis used to determine the nature of the interaction of the two drugs. Flow cytometry and western blot were employed to demonstrate drug effects on cell cycle distribution and phosph-Aktser473 expression. Radiolabelled ([methyl-3H]) Choline uptake was measured in control and drug-treated cells to determine the modulatory effects of the drugs on choline incorporation. The two drugs acted synergistically to inhibit the growth rate of each cancer cell line. Flow cytometry demonstrated G0/G1 blockade with MK2206 and AZD8055 which was greater when cells were treated with both drugs. The incorporation of [methyl-3H] choline was found be decreased to a greater extent in cells treated with both drugs compared with cells treated with either drug alone. Conclusions Pan-mTOR and pan-Akt inhibition may be highly effective in cancer treatment and measuring changes in choline uptake could be useful in detecting efficacious drug combinations.
Highlights
Cancer growth and development are frequently associated with upregulation of intracellular signalling pathways, including the PI3K/Akt/mTOR pathway, which drives cellular proliferation resulting in the uncontrolled cell growth that defines cancer [1]
In view of the central role of PI3K/Akt/mTOR in cancer cell proliferation in many tumour types there is much interest in oncology in the use of inhibitors targeting components of this pathway [3]. The use of such inhibitors has been shown to decrease cancer cell proliferation and survival, their efficacy as single-agents is poor [4] due to activation of feedback mechanisms [5, 6] so attenuating the clinical benefit from the drug. Their use in combination treatments has been recommended [7] and it has been shown that drug combinations, including the dual use of Akt and mTOR inhibitors, are more efficacious than single drug regimens as they bring about a more complete pathway inhibition [8]
We have demonstrated that treatment of breast cancer and colorectal cancer cells with a pan-mTOR inhibitor synergistically increases the growthinhibitory action of the Akt-inhibitor MK2206 which corresponded with changes in the incorporation of radiolabelled choline
Summary
Cancer growth and development are frequently associated with upregulation of intracellular signalling pathways, including the PI3K/Akt/mTOR pathway, which drives cellular proliferation resulting in the uncontrolled cell growth that defines cancer [1]. The use of such inhibitors has been shown to decrease cancer cell proliferation and survival, their efficacy as single-agents is poor [4] due to activation of feedback mechanisms [5, 6] so attenuating the clinical benefit from the drug. Their use in combination treatments has been recommended [7] and it has been shown that drug combinations, including the dual use of Akt and mTOR inhibitors, are more efficacious than single drug regimens as they bring about a more complete pathway inhibition [8]. Drugs targeting the PI3K/ Akt/mTOR pathway are associated with unpleasant and potentially dangerous side effects including nausea, diarrhoea and hyperglycaemia [9] which may be enhanced in drug combinations [8]
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