KU-60019 radiosensitized U87 glioma cells. Normal fibroblasts were also radiosensitized, whereas ATM-deficient A-T fibroblasts were not further sensitized under the tested conditions, supporting ATM dependence without demonstrating tumor specificity. [1]
In colorectal cancer cells, KU-60019 combined with the Chk1 inhibitor LY2606368 reduced viability more effectively than the individual agents at tested concentrations. Normal SF-TY fibroblast viability also decreased with combination treatment, limiting claims of cancer-cell selectivity. [2]
Inhibition of radiation-induced ATM signaling
Cell-based study · Human U87 and U1242 glioma cells [1]
KU-60019 suppressed radiation-induced p53 (S15) and CHK2 (T68) phosphorylation. H2AX phosphorylation was less completely inhibited; this is a cellular signaling endpoint, not an enzyme IC50 assay.
Literature-Reported Treatment Conditions
- Treatment concentration
- U87: 0, 1, 3 or 10 μM; U1242: 3 μM
- Exposure duration
- U87: 1 h pretreatment, harvest 1 h after irradiation; U1242: harvest 5, 15, 30 or 60 min after irradiation
- Method
- Western blotting with β-actin normalization
- Co-treatment
- U87: 10 Gy ionizing radiation; U1242: 5 Gy
- Reported limits
- U1242 pretreatment interval is not specified in Figure 2C caption; vehicle and replicate count for these blots are not stated in the acquired selected methods/caption.
Methods and source locations
Compound reported in the paper: KU-60019
Evidence concerns the named KU-60019 reagent in these models; no salt conversion or Adooq lot equivalence is established.
Figure 2A,C and caption; Results: KU-60019 is a more potent inhibitor of the ATM kinase than KU-55933; Materials and Methods: Chemical synthesis, Reagents, Western blotting
Radiation-induced p53 S15 phosphorylation in U87 cells: Western blot; unirradiated, radiation-alone and KU-55933 comparison lanes; β-actin control — Figure 2A and Results paragraph
Radiation-induced CHK2 T68 and H2AX S139 phosphorylation in U1242 cells: Western blot, drug-free irradiation controls and β-actin normalization — Figure 2C and caption; accompanying Results
Radiosensitization with normal and ATM-deficient controls
Cell-based study · U87 human glioma cells; normal and A-T human fibroblasts [1]
KU-60019 radiosensitized U87 cells. Normal fibroblasts also responded, whereas A-T fibroblasts were not further radiosensitized, supporting ATM dependence without establishing tumor specificity.
Literature-Reported Treatment Conditions
- Treatment concentration
- U87: 1 or 10 μM KU-60019; normal/A-T fibroblasts: 10 μM
- Exposure duration
- 1 h before irradiation; drug removed 16 h after irradiation; survival assessed 7 days after irradiation
- Method
- Trypan blue/flow-cytometry survival assay
- Co-treatment
- Ionizing radiation; U87 comparator KU-55933 at 10 μM
- Reported limits
- Detailed supplemental flow-cytometry protocol was not retrieved; DER is preserved as reported without assigning an unverified survival-fraction definition. Vehicle not specified in acquired selected methods/caption.
Methods and source locations
Compound reported in the paper: KU-60019
Evidence concerns the named KU-60019 reagent in these models; no salt conversion or Adooq lot equivalence is established.
Figure 3A,D and caption; Results: KU-60019 is a more potent radiosensitizer than KU-55933 and KU-60019 specifically targets the ATM kinase to radiosensitize cells; Cell survival methods
U87 radiosensitization: Trypan blue/FACS at day 7; surviving fraction relative to nonirradiated control; caption: SEM, n = 3, replicate type unspecified — Figure 3A, printed DER labels and caption; Results radiosensitization paragraph
Normal versus A-T fibroblast radiosurvival: Trypan blue/FACS at day 7; drug-free irradiation controls; caption: SEM, n = 3 — Figure 3D and caption; Results specificity paragraph
KU-60019 and LY2606368 combination viability response
Cell-based study · Human colorectal cancer HCT116, DLD-1 and HT29 cells; SF-TY normal human fibroblasts [2]
The combination decreased colorectal-cell viability at lower concentrations than single agents and produced CI values interpreted as synergy. Normal-fibroblast viability also decreased; the result does not demonstrate normal-cell sparing.
Literature-Reported Treatment Conditions
- Treatment concentration
- Figure 1B: KU-60019 concentration series with LY2606368 fixed at 0.03 μM for HCT116/DLD-1 or 3 nM for HT29. Figure 1C: LY2606368 series with KU-60019 fixed at 10 μM for HCT116/DLD-1 or 3 μM for HT29. SF-TY: corresponding series with fixed LY2606368 0.03 μM or KU-60019 10 μM.
- Exposure duration
- 48 h drug treatment; WST-8 incubation 1 h
- Method
- WST-8 cell proliferation/viability assay, absorbance 450 nm
- Vehicle
- DMSO stock solutions diluted into culture medium; final DMSO concentration not specified
- Co-treatment
- LY2606368 (Chk1 inhibitor)
- Reported limits
- Exact CI and IC50 tables in SI not retrieved; no decimal values reconstructed from curves. Caption n = 5 does not specify biological versus technical replicates; untreated control is explicit, matched final-DMSO control is not described.
Methods and source locations
Compound reported in the paper: KU-60019
Evidence concerns the named KU-60019 reagent in these models; no salt conversion or Adooq lot equivalence is established.
Figure 1A–E and caption; Methods 2.1–2.2; Results 3.1–3.2
Colorectal cancer-cell viability and combination interaction: WST-8, untreated viability defined as 1; mean ± SE, n = 5; Tukey multiple comparison test, p < 0.05 symbols for indicated comparisons — Figure 1A–D and caption; Results 3.1–3.2; Methods 2.2
Normal SF-TY fibroblast viability: WST-8 after 48 h; untreated viability defined as 1; mean ± SE, n = 5 — Figure 1E and caption; Results 3.2 final paragraph
