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. 2021 May 20;10(5):1261. doi: 10.3390/cells10051261

Figure 5.

Figure 5

Radiotherapy and schedule-dependent PF-04691502 therapy in NET cells. (A) QGP-1 (top) and BON (bottom) cells were seeded in a 6-well plate at 800,000 cells/2 mL density. Cells were then exposed to either control (0 Gy), 2, 4, or 8 Gy X-ray radiation. Protein lysates were collected at 24, 48, 72, and 96 h after radiation exposure and analyzed for cleaved-PARP expression by immunoblotting. β-actin was used as a loading control. (B) QGP-1 (top) and BON (bottom) cells were seeded in 6-well plate at 800,000 cells/2 mL density and exposed to 2 Gy X-ray radiation. Cells were incubated for either 48, 72, or 96 h and then treated with PF-04691502 at 500 or 1000 nM for 24 h prior to protein lysates collection. Cleaved-PARP expression was determined by immunoblotting analysis. β-actin was used as a loading control. (C) QGP-1 and BON cells were seeded at 1000 cells/100 uL density in 96-well plates and exposed to 2 Gy X-ray radiation. Cells were incubated for 96 h and treated with PF-04691502 at 500 nM for 24 h prior to lysate collection and analysis as described. DNA fragmentation and cell death induced proportional colorimetric change, which was measured by absorbance at 405 nm (n = 6). * denotes p-value < 0.01. (D) Proposed schematic diagram for schedule-dependent PI3K/mTOR inhibition after radiotherapy in NET patients.