Article Figures & Data
Figures
- Fig. 1.
The effect of SCH66336 on HNSCC cells. A subset of HNSCC cells was treated with 0.1% DMSO or SCH66336 (SCH; 5 μmol/L) for 48 h and analyzed. Percentage of cells in specific phases of the cell cycle (sub-G1, G0-G1, S, and G2-M) by flow cytometric analysis (A) and a flow cytometry–based terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling assay (B).
- Fig. 2.
The effect of SCH66336 on the expression of survivin. A, Western blot analysis in a normal human oral keratinocytes (NHOK) and in a subset of HNSCC cells. B, flow cytometry–based terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling assay (top) and Western blot analysis (bottom) in TR146, UMSCC38, and SqCC/Y1 cells incubated for 48 h in medium with 0.1% FBS and in the presence or absence of 5 μmol/L SCH66336. C, Western blot analysis in a subset of HNSCC cells, which were incubated for 48 h in 10% FBS medium with or without 5 μmol/L SCH66336. D, cell proliferation (72 h) and Western blot analysis (48 h) of H460, 226Br, and 226B NSCLC cell lines, which were treated with 5 μmol/L SCH66336.
- Fig. 3.
Survivin mediates resistance to SCH66336 in cancer cells. Flow cytometry–based terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling assay (A) and Western blotting (B) in TR146 and UMSCC38 cells after transfection with control siRNA (si-scr) or survivin siRNA (si-survivin) followed by treatment with 5 μmol/L SCH66336 for 48 h. C, Western blotting in SqCC/Y1, UMSCC10B, and SqCC35 cells infected with adenoviral vector expressing survivin (Ad-survivin) or empty vector (Ad-EV) followed by 5 μmol/L SCH66336 for 48 h. Ac-caspase-3, active caspase-3.
- Fig. 4.
SCH66336 induced survivin expression through the IGF-IR pathway. A, reverse transcription-PCR analysis of survivin in TR146, UMSCC38, and SqCC/Y1 cells after treatment with 5 μmol/L SCH66336 for 48 h. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the control. B, Western blotting in a subset of HNSCC cells incubated for 48 h in the medium containing 10% FBS with or without 5 μmol/L SCH66336. C, proliferation of TR146 and UMSCC38 cells after adenoviral vector (Ad-EV) or adenoviral vector expressed truncated α-subunit of IGF-IR (Ad-dnIGF-IR) into the medium after treatment with 5 μmol/L SCH66336 for 48 h. Cell proliferation (72 h; D) and Western blotting (48 h; D, bottom) in TR146 and UMSCC38 cells after 5 μmol/L SCH66336 and IGF-IR tyrosine kinase inhibitor AG1024 treatment. **, P < 0.01; ***, P < 0.001, compared with indicated control.
- Fig. 5.
Inhibition of Akt/mTOR pathway enhances the apoptotic effect of SCH66336 in HNSCC cells. Western blot analysis (48 h; A) and cell proliferation (72 h; B) in TR146 cells after treatment with 5 μmol/L SCH66336 plus the PI3K inhibitor LY194002 (LY) or 5 μmol/L SCH66336 plus the MAPK/extracellular signal-regulated kinase kinase inhibitor U0126. Ac-caspase-3, cleaved caspase-3. Western blot analysis (48 h; C) and cell proliferation (72 h; D) of TR146 cells after treatment with 5 μmol/L SCH66336 plus 100 nmol/L mTOR inhibitor rapamycin (Rapa). ***, P < 0.001, compared with indicated control.
Volume 14, Issue 5
