Cellular responses depend on the model and conditions. In SCC-9 cells, 3 μM GSK1059615 did not significantly induce apoptosis at 24–72 h despite an active gemcitabine positive control. In HT-p21 cells, proliferation inhibition had a reported IC50 >300 nM and was reversible after drug removal in the tested washout experiment. [2,3]
Biochemical inhibition of PI3Kα
Biochemical assay · PI3Kα p110α/p85α biochemical assay [1]
Compound 2 inhibited PI3Kα with a reported IC50 of 2 nM. This is an assay endpoint, not a treatment concentration. Compound 1 is GSK2126458 and its potency and animal experiments are not transferred.
Literature-Reported Treatment Conditions
- Method
- Continuous-read time-resolved fluorescence resonance energy transfer displacement assay
- Reported limits
- Full assay conditions, control calibration, ATP conditions and replicate statistics in linked SI were not retrieved; no isoform panel or mTOR IC50 is adopted.
Methods and source locations
Compound reported in the paper: GSK1059615; Compound label in the paper: Compound 2
Evidence concerns the named research compound in the reported model; no validation of the current Adooq lot, formulation equivalence or clinical benefit.
Figure 1; Table 1, compound 2 row; PI3Kα assay paragraph and biochemical comparison paragraph
Inhibition of PI3Kα: TR-FRET displacement assay; original reported value, not curve-derived — Table 1, compound 2 row; biochemical comparison paragraph
PI3K–AKT–mTOR signaling in HNSCC cells
Cell-based study · SCC-9 cells, primary oral cavity carcinoma OCC1 cells, and oral epithelial Oepi1 cells [2]
Phosphorylation of PI3K p85, AKT, mTOR and S6K1 decreased in SCC-9 and OCC1 cells; ERK1/2 phosphorylation was not altered. Oepi1 had low basal PI3K–AKT–mTOR pathway phosphorylation, limiting a general normal-cell selectivity inference.
Literature-Reported Treatment Conditions
- Treatment concentration
- 3 μM
- Exposure duration
- 2 h
- Method
- Cell-lysate protein phosphorylation analysis versus untreated control
- Reported limits
- Solvent concentration is not specified for this signaling experiment. Figure legend reports three repeats; biological versus technical repeat identity is not defined.
Methods and source locations
Compound reported in the paper: GSK1059615; Compound label in the paper: GSK1059615
Evidence concerns the named research compound in the reported model; no validation of the current Adooq lot, formulation equivalence or clinical benefit.
Results: GSK1059615 blocks PI3K-AKT-mTOR activation; Figure 3A–C and caption; Chemicals and reagents; cell culture sections
PI3K p85 Tyr458, AKT Ser473, mTOR Ser2448 and S6K1 Thr389 phosphorylation; ERK1/2 control: Figure 3A–C; three repeats; bars mean ± SD by general figure convention; no numerical bar heights extracted — Figure 3A–C, caption and corresponding Results
Negative apoptosis findings with an active positive control
Cell-based study · SCC-9 human HNSCC cells [2]
GSK1059615 did not significantly increase apoptosis or obvious caspase cleavage under these conditions. Gemcitabine served as an active positive control. This limits an unqualified apoptosis-inducer description.
Literature-Reported Treatment Conditions
- Treatment concentration
- 3 μM
- Exposure duration
- 24, 48 and 72 h
- Method
- Annexin V FACS, TUNEL, histone–DNA apoptosis ELISA, and caspase-3/-9 cleavage analysis; untreated control and gemcitabine 500 nM for 48 h
- Reported limits
- Solvent fraction not specified for this experiment; independence of assay replicates not defined.
Methods and source locations
Compound reported in the paper: GSK1059615; Compound label in the paper: GSK1059615
Evidence concerns the named research compound in the reported model; no validation of the current Adooq lot, formulation equivalence or clinical benefit.
Results: GSK1059615 fails to provoke apoptosis; Figure 4A–D and caption; apoptosis assay methods; Chemicals and reagents
Apoptotic cells and cleaved caspase-3/-9: n=5 for each assay; experiments repeated three times; general bars mean ± SD. Gemcitabine 500 nM, 48 h increased apoptosis. No exact plotted percentages extracted. — Figure 4A–D and caption; corresponding Results
Reversible growth inhibition in HT-p21 cells
Cell-based study · HT-p21 cells derived from human HT1080 fibrosarcoma [3]
GSK1059615 inhibited proliferation, with reported IC50 >300 nM. Cells resumed proliferation after drug removal in the tested washout experiment, supporting reversible cytostasis in this model.
Literature-Reported Treatment Conditions
- Treatment concentration
- Serial dilutions; GSK1059615 plotted through 300 nM in Figure 1A–B
- Exposure duration
- 4-day exposure; after replating in drug-free medium, a further 4 days for Figure 1B
- Method
- Cell counting in triplicate; washout/replating follow-up versus untreated control
- Vehicle
- DMSO stock; final DMSO fraction not specified
- Reported limits
- No salt, catalog/lot, final solvent fraction or explicit biological replicate identity supplied. No maximum gerosuppression concentration is adopted from unviewed SI.
Methods and source locations
Compound reported in the paper: GSK1059615; Compound label in the paper: GSK1059615
Evidence concerns the named research compound in the reported model; no validation of the current Adooq lot, formulation equivalence or clinical benefit.
Results opening paragraph; Figure 1A–B and caption; Cell lines and reagents
Proliferation measured by cell number: 4 days; triplicate counts; mean ± SD; value explicitly stated in Results rather than fitted from plot — Results opening paragraph; Figure 1A and caption
Cell proliferation after drug removal: 4 days exposure then counted/replated at 1000 cells/well; 4 days in drug-free medium; not the 7-day IPTG gerosuppression design — Figure 1B and caption; Results opening paragraph
