Article Figures & Data
Figures
- Figure 1.
Augmentation of TRAIL-mediated caspase activity and c-FLIPL down-regulation by chemical inhibition of CK2 in prostate cancer cells. Caspase activity was assessed by spectrofluorimetric assay, as described in Materials and Methods. A, ALVA-41 cells (0.25 × 106) treated with TBB (40 μmol/L) or TRAIL alone (2.0 ng/mL) for 24 hours and after pretreatment with 40 μmol/L TBB for 3 hours and subsequent addition of TRAIL for 21 hours. B, PC-3 cells (0.25 × 106) treated with TBB (60 μmol/L) or TRAIL alone (10 ng/mL) for 24 hours and after pretreatment with 60 μmol/L TBB for 3 hours and subsequent addition of TRAIL for 21 hours. C, processing of caspase-3 and caspase-8, and c-FLIPL expression in PC-3 cells was determined by immunoblot analysis. Lane a, untreated control; lane b, TRAIL (10 ng/mL); lane c, TBB (60 μmol/L); lane d, TBB (60 μmol/L) plus TRAIL (10 ng/mL).
- Figure 2.
Sensitization of TRAIL-induced apoptosis in prostate cancer cells by treatment with TBB. ALVA-41 and PC-3 cells (0.25 × 106) were treated with TRAIL in the presence or absence of TBB as in Fig. 1. A, cell death in ALVA-41 and PC-3 cells was assessed by WST-1 assay. B, DNA fragmentation in ALVA-41 cells was measured by propidium iodide staining. C, lysates from PC-3 and ALVA-41 cells treated under the same conditions as above were subjected to immunoblot analysis using anti-lamin A as described in Materials and Methods. Lane a, untreated control; lane b, TRAIL; lane c, TBB; lane d, TBB plus TRAIL.
- Figure 3.
Effect of TBB on TRAIL-mediated expression of Bax, Bcl-2, Bcl-XL, and release of cytochrome c in prostate cancer cells. Lysates prepared from ALVA-41 and PC-3 (2.5 × 106) cells treated with desired concentrations of TBB with or without TRAIL as in Fig. 1 were subjected to immunoblot analysis for anti-Bax, anti-Bcl-2, anti-Bcl-XL, anti-cytochrome c, and anti-actin. Lane a, untreated control; lane b, TRAIL; lane c, TBB; lane d, TBB plus TRAIL.
- Figure 4.
Effect of overexpression of CK2 on TRAIL-induced caspase activity in prostatic cancer cells. Caspase activity was determined by spectrofluorimetric assay as described in Materials and Methods. A, ALVA-41 cells (0.25 × 106) were transfected with pcDNA6-CK2α (2 μg/mL) using DOTAP transfection reagent for 24 hours followed by treatment with 10 ng/mL of TRAIL for 24 hours. B, PC-3 cells (0.25 × 106) were transfected with pcDNA6-CK2α (2 μg/mL) for 24 hours, followed by 25 ng/mL of TRAIL for 24 hours. C, representative immunoblot analysis of processing of caspase-3, caspase-8, and c-FLIPL is shown for PC-3 cells. Lane a, untreated control; lane b, TRAIL (25 ng/mL); lane c, pcDNA6-CK2α (2 μg/mL), and lane d, pcDNA6-CK2α (2 μg/mL) plus TRAIL (25 ng/mL).
- Figure 5.
Effect of overexpression of CK2 on TRAIL-induced apoptosis in prostate cancer cells. A, ALVA-41 and PC-3 (0.25 × 106) cells were transfected with pcDNA6-CK2α (2 μg/mL) using DOTAP for 24 hours followed by treatment with 10 ng/mL TRAIL for ALVA-41 and 25 ng/mL TRAIL for PC-3 cells. Cell death was assessed by WST-1 assay. B, PC-3 (0.25 × 106) cells were transfected with pcDNA6-CK2α (2 μg/mL) using DOTAP for 24 hours followed by TRAIL (25 ng/mL) for 24 hours. DNA fragmentation was assayed by propidium iodide staining. C, lysates from treated cells were subjected to immunoblot analysis for lamin A cleavage. Lane a, untreated control; lane b, TRAIL; lane c, pcDNA6-CK2α; and lane d, pcDNA6-CK2α plus TRAIL.
- Figure 6.
Effect of overexpression of CK2 on mitochondrial engagement by TRAIL in prostate cancer cells. ALVA-41 and PC-3 cells (2.5 × 106) were transfected in T25 flasks with pcDNA6-CK2α (2.0 μg/mL) for 24 hours using DOTAP, followed by treatment with 10 ng/mL TRAIL for ALVA-41 cells and 25 ng/mL for PC-3 cells for 24 hours. Lysates of the treated cells were subjected to immunoblot analysis using anti-Bax, anti-Bcl-2, anti-Bcl-XL, anti-cytochrome c, and anti-actin antibodies. Lane a, untreated control; lane b, TRAIL; lane c, pcDNA6-CK2α; and lane d, pcDNA6-CK2α plus TRAIL.
Volume 66, Issue 4
