Abstract
BackgroundThe nudix family member enzyme MutT homologue-1 (MTH1) hydrolyses the oxidized nucleotides 8-oxo-dGTP and 2-hydroxy-dATP and thus prevents the incorporation of damaged nucleotides into nuclear and mitochondrial DNA. Therefore MTH1 was proposed to protect cancer cells from oxidative DNA lesions and subsequent cell death. We investigated whether the bona fide MTH1 inhibitor TH588 affects responses of cultured colorectal tumor cells to ionizing radiation (IR) in normoxia and in moderate or severe hypoxia.MethodsTH588 was tested in cell viability and survival assays (tetrazolium dye (MTT), propidium iodide staining, caspase-3 activity, and colony formation assays (CFA)) in colorectal carcinoma cells (HCT116 and SW480) in combination with IR in normoxia and in hypoxia. Additionally, MTH1 was targeted by lentiviral shRNA expression. Human umbilical vein endothelial cells (HUVEC) were assessed in MTT assays.ResultsIn all cell lines tested, TH588 dose-dependently impaired cell survival. In CFAs, TH588 and IR effects on carcinoma cells were additive in normoxia and in hypoxia. Using 3 different shRNAs, the lentiviral approach was detrimental to SW480, but not to HCT116.ConclusionsTH588 has cytotoxic effects on transformed and untransformed cells and synergizes with IR in normoxia and in hypoxia. TH588 toxicity is not fully explained by MTH1 inhibition as HCT116 were unaffected by lentiviral suppression of MTH1 expression. TH588 should be explored further because it has radiosensitizing effects in hypoxia.
Highlights
The nudix family member enzyme MutT homologue-1 (MTH1) hydrolyses the oxidized nucleotides 8-oxo-dGTP and 2-hydroxy-dATP and prevents the incorporation of damaged nucleotides into nuclear and mitochondrial DNA
Because Human umbilical vein endothelial cells (HUVEC) are widely used as a model system for untransformed human cells, we tested their response to MTH1 inhibition
After plating 8.000 cells viability was significantly reduced to 40% of the control at a concentration of 5 μM TH588 (Fig. 1a) and very similar to the cancer cell lines tested
Summary
The nudix family member enzyme MutT homologue-1 (MTH1) hydrolyses the oxidized nucleotides 8-oxo-dGTP and 2-hydroxy-dATP and prevents the incorporation of damaged nucleotides into nuclear and mitochondrial DNA. By hydrolysis of 8-oxo-dGTP, MTH1 prevents incorporation of 8oxoG into DNA [4]. Crizotinib, a drug which is in clinical use and regarded as a tyrosin kinase inhibitor, has been reported to inhibit MTH1 [11, 12]. These compounds including TH588 bind to the active site of MTH1 and prevent access of 8-oxo-dGTP. In the same publication toxicity is proposed to be limited to tumor cells
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