RAD001 combination profiling and signaling in NCI-H446 cells
Cell-based study · Human NCI-H446 small cell lung cancer cells [1]
RAD001 combinations with BYL719 or MK-2206 showed ZIP synergy in a 72-h cell viability assay. CAL-101/RAD001 was additive under the authors’ >10 synergy criterion. Companion immunoblots included single-agent and combination controls; these data do not establish single-agent apoptosis or animal efficacy of RAD001.
| Endpoint | Reported result | Reference |
|---|---|---|
| RAD001 plus BYL719 interaction in cell viability | ZIP synergy score 12.889 | [1] |
| RAD001 plus MK-2206 interaction in cell viability | ZIP synergy score 10.688 | [1] |
| CAL-101 plus RAD001 interaction in cell viability | ZIP synergy score 5.878 | [1] |
| Single-agent RAD001 signaling response | qualitative immunoblot Reduced p-P70S6K(T389), with retained p-4EBP1(T37/T46) signal and increased p-AKT signals relative to DMSO at 1 µM RAD001 | [1] |
Literature-Reported Treatment Conditions
- Treatment concentration
- RAD001 matrix: 0–1000 nM; BYL719 or MK-2206 matrix: 0–10 µM (Fig. 2C). Single-agent signaling: 10, 100 and 1000 nM RAD001 (Fig. 2B). Fig. 2E single/combination treatments: 1.0 µM per indicated drug.
- Exposure duration
- 72 h for viability matrices; 24 h for Fig. 2B immunoblots.
- Method
- Cell treatment followed by PrestoBlue fluorescence viability or Western blotting.
- Vehicle
- DMSO lane and mock controls in Fig. 2B; viability normalized to untreated cells. Vehicle percentage for viability not specified in the read methods.
- Co-treatment
- BYL719 (alpelisib), MK-2206 or CAL-101 in distinct combinations with RAD001; effects are not assigned to RAD001 alone.
- Reported limits
- No current-lot analytical validation; exact viability vehicle percentage and independent matrix repeat count not specified. Fig. 2E duration is not stated in its caption and is not assumed from Fig. 2B.
Methods and source locations
Compound reported in the paper: Everolimus; Compound label in the paper: RAD001
Evidence applies to the named tested material and model; it does not validate a current Adooq lot or establish clinical efficacy.
Methods: Test compounds, Cell viability, Immunoblotting; Results: Synergistic effects of PI3Kα and mTOR; Fig. 2B–E and caption
RAD001 plus BYL719 interaction in cell viability: PrestoBlue viability normalized to untreated cells; SynergyFinder ZIP model. Eight-point titrations for general viability methods were performed in triplicate; independent repeat count for the synergy matrix is not specified in Fig. 2. — Fig. 2C–D; Methods: Cell viability
RAD001 plus MK-2206 interaction in cell viability: Same combination profiling design. — Fig. 2C–D
CAL-101 plus RAD001 interaction in cell viability: ZIP; authors classify as additive, using >10 as the synergy criterion. — Fig. 2D; Results: Synergistic effects
Single-agent RAD001 signaling response: 24-h Western blot with total-protein, β-ACTIN and GAPDH controls; qualitative assessment, no densitometry estimated. — Fig. 2B and caption