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Nexavar/Stivarga and viagra interact to kill tumor cells.

Tavallai M, Hamed HA, Roberts JL, Cruickshanks N, Chuckalovcak J, Poklepovic A, Booth L, Dent P - J. Cell. Physiol. (2015)

Bottom Line: PDE5 and PDGFRα/β were over-expressed in liver tumors compared to normal liver tissue.Knock down of PDE5 or of PDGFRα/β recapitulated the effects of the individual drugs.The drug combination also reduced mTOR protein expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, Virginia.

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Related in: MedlinePlus

Treatment of HT29 tumors with regorafenib, sildenafil and FTY720 alters the cytokine expression levels in mouse plasma. (A–G) HT29 tumors (~30 mm3) were formed in the flanks of athymic mice. Aliquots (~75 μl) of mouse blood were obtained in a heparin/EDTA coated Eppendorf tube. Animals were then treated PO with vehicle diluent (cremophore); sildenafil (5 mg/kg) and regorafenib (25 mg/kg); FTY720 (0.05 mg/kg) or the drugs in combination as indicated for 7 days. Data in part A are vehicle control and [regorafenib + sildenafil] tumor data. After 7 days aliquots of mouse blood were again obtained (~75 μl). Clarified mouse plasma free of cells was then subjected to multiplex assays in a Bio-Rad MAGPIX system to define the expression of the noted cytokines in each part before and following treatment (n = 2 studies, eight animals per group ±SEM). *P < 0.05 greater than Day 0 pre-treatment value; **P < 0.05 greater than Day 7 vehicle control; #P < 0.05 less than Day 0 pre-treatment value; ##P < 0.05 less than Day 7 vehicle value; ¶P < 0.05 less than Day 7 regorafenib treatment value. (H) HT29 and HCT116 tumor cells in vitro were treated with vehicle control; [regorafenib (0.5 μM) + FTY720 (50 nM)]; the TGF β receptor inhibitor LY2157299 (0.2, 0.6 μM) or the drugs in combination as indicted for 9 h. After 9 h cell viability was determined in a Hermes WiScan instrument using a live/dead assay (n = 3, ±SEM). *P < 0.05 greater than REGO + FTY value. (I–L) HT29 tumors isolated from vehicle control treated or (regorafenib + sildenafil) treated tumors at the time of animal nadir were subjected to multiplex assays in a Bio-Rad MAGPIX analyzer to determine the expression of cytokines in plasma and the phosphorylation of the indicated signal transduction proteins (n = 8, ±SEM). ~P < 0.05 greater than vehicle control treated tumor value; #P < 0.05 less than vehicle control treated tumor value. (M) HuH7 cells were treated with vehicle control; regorafenib (0.5 μM) and sildenafil (2 μM); BGJ398 (1 μM); Lapatinib (1 μM); MK2206 (1 μM) or in the combinations indicated in the part. Cells were examined 12 h after treatment using a live/dead assay in a Hermes WiScan instrument. Red/yellow cells = dead; green cells = alive (n = 3, ±SEM). *P < 0.05 greater than corresponding value in (regorafenib + sildenafil) treatment alone.
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Figure 10: Treatment of HT29 tumors with regorafenib, sildenafil and FTY720 alters the cytokine expression levels in mouse plasma. (A–G) HT29 tumors (~30 mm3) were formed in the flanks of athymic mice. Aliquots (~75 μl) of mouse blood were obtained in a heparin/EDTA coated Eppendorf tube. Animals were then treated PO with vehicle diluent (cremophore); sildenafil (5 mg/kg) and regorafenib (25 mg/kg); FTY720 (0.05 mg/kg) or the drugs in combination as indicated for 7 days. Data in part A are vehicle control and [regorafenib + sildenafil] tumor data. After 7 days aliquots of mouse blood were again obtained (~75 μl). Clarified mouse plasma free of cells was then subjected to multiplex assays in a Bio-Rad MAGPIX system to define the expression of the noted cytokines in each part before and following treatment (n = 2 studies, eight animals per group ±SEM). *P < 0.05 greater than Day 0 pre-treatment value; **P < 0.05 greater than Day 7 vehicle control; #P < 0.05 less than Day 0 pre-treatment value; ##P < 0.05 less than Day 7 vehicle value; ¶P < 0.05 less than Day 7 regorafenib treatment value. (H) HT29 and HCT116 tumor cells in vitro were treated with vehicle control; [regorafenib (0.5 μM) + FTY720 (50 nM)]; the TGF β receptor inhibitor LY2157299 (0.2, 0.6 μM) or the drugs in combination as indicted for 9 h. After 9 h cell viability was determined in a Hermes WiScan instrument using a live/dead assay (n = 3, ±SEM). *P < 0.05 greater than REGO + FTY value. (I–L) HT29 tumors isolated from vehicle control treated or (regorafenib + sildenafil) treated tumors at the time of animal nadir were subjected to multiplex assays in a Bio-Rad MAGPIX analyzer to determine the expression of cytokines in plasma and the phosphorylation of the indicated signal transduction proteins (n = 8, ±SEM). ~P < 0.05 greater than vehicle control treated tumor value; #P < 0.05 less than vehicle control treated tumor value. (M) HuH7 cells were treated with vehicle control; regorafenib (0.5 μM) and sildenafil (2 μM); BGJ398 (1 μM); Lapatinib (1 μM); MK2206 (1 μM) or in the combinations indicated in the part. Cells were examined 12 h after treatment using a live/dead assay in a Hermes WiScan instrument. Red/yellow cells = dead; green cells = alive (n = 3, ±SEM). *P < 0.05 greater than corresponding value in (regorafenib + sildenafil) treatment alone.

Mentions: Based on our unexpected findings in Figure 9, we determined using a Bio-Rad MAGPIX multiplex system the expression levels of human cytokines in mouse plasma and the expression and activity of signal transduction proteins within the established tumor itself. Regorafenib and sildenafil exposure for 7 days decreased the plasma levels of bFGF and GM-CSF but significantly increased the expression of PDGFbb (Fig. 10A). In agreement with data in Figure 8, treatment of tumors for 7 days with FTY720 increased plasma levels of FAS-L (Fig. 10B). For a number of cytokines, e.g., endoglin, IL6, VEGF-A/C/D, endpoietin 2, EGF, their plasma level declined in vehicle control treated tumors over 7 days of growth which was associated with a mean of approximately sevenfold increase in tumor volume (Fig. 10B–F). In this respect, treatment of tumors with FTY720 or with regorafenib and FTY720 prevented the decline in plasma cytokine levels that were observed in mice with vehicle control treated tumors at day 7 (Fig. 10B–G). For tumors exposed to regorafenib and sildenafil, the expression of pro-growth/pro-angiogenic/pro-invasion cytokines was either unchanged compared to vehicle control treated tumors or was significantly reduced (Fig. 10C–G).


Nexavar/Stivarga and viagra interact to kill tumor cells.

Tavallai M, Hamed HA, Roberts JL, Cruickshanks N, Chuckalovcak J, Poklepovic A, Booth L, Dent P - J. Cell. Physiol. (2015)

Treatment of HT29 tumors with regorafenib, sildenafil and FTY720 alters the cytokine expression levels in mouse plasma. (A–G) HT29 tumors (~30 mm3) were formed in the flanks of athymic mice. Aliquots (~75 μl) of mouse blood were obtained in a heparin/EDTA coated Eppendorf tube. Animals were then treated PO with vehicle diluent (cremophore); sildenafil (5 mg/kg) and regorafenib (25 mg/kg); FTY720 (0.05 mg/kg) or the drugs in combination as indicated for 7 days. Data in part A are vehicle control and [regorafenib + sildenafil] tumor data. After 7 days aliquots of mouse blood were again obtained (~75 μl). Clarified mouse plasma free of cells was then subjected to multiplex assays in a Bio-Rad MAGPIX system to define the expression of the noted cytokines in each part before and following treatment (n = 2 studies, eight animals per group ±SEM). *P < 0.05 greater than Day 0 pre-treatment value; **P < 0.05 greater than Day 7 vehicle control; #P < 0.05 less than Day 0 pre-treatment value; ##P < 0.05 less than Day 7 vehicle value; ¶P < 0.05 less than Day 7 regorafenib treatment value. (H) HT29 and HCT116 tumor cells in vitro were treated with vehicle control; [regorafenib (0.5 μM) + FTY720 (50 nM)]; the TGF β receptor inhibitor LY2157299 (0.2, 0.6 μM) or the drugs in combination as indicted for 9 h. After 9 h cell viability was determined in a Hermes WiScan instrument using a live/dead assay (n = 3, ±SEM). *P < 0.05 greater than REGO + FTY value. (I–L) HT29 tumors isolated from vehicle control treated or (regorafenib + sildenafil) treated tumors at the time of animal nadir were subjected to multiplex assays in a Bio-Rad MAGPIX analyzer to determine the expression of cytokines in plasma and the phosphorylation of the indicated signal transduction proteins (n = 8, ±SEM). ~P < 0.05 greater than vehicle control treated tumor value; #P < 0.05 less than vehicle control treated tumor value. (M) HuH7 cells were treated with vehicle control; regorafenib (0.5 μM) and sildenafil (2 μM); BGJ398 (1 μM); Lapatinib (1 μM); MK2206 (1 μM) or in the combinations indicated in the part. Cells were examined 12 h after treatment using a live/dead assay in a Hermes WiScan instrument. Red/yellow cells = dead; green cells = alive (n = 3, ±SEM). *P < 0.05 greater than corresponding value in (regorafenib + sildenafil) treatment alone.
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Figure 10: Treatment of HT29 tumors with regorafenib, sildenafil and FTY720 alters the cytokine expression levels in mouse plasma. (A–G) HT29 tumors (~30 mm3) were formed in the flanks of athymic mice. Aliquots (~75 μl) of mouse blood were obtained in a heparin/EDTA coated Eppendorf tube. Animals were then treated PO with vehicle diluent (cremophore); sildenafil (5 mg/kg) and regorafenib (25 mg/kg); FTY720 (0.05 mg/kg) or the drugs in combination as indicated for 7 days. Data in part A are vehicle control and [regorafenib + sildenafil] tumor data. After 7 days aliquots of mouse blood were again obtained (~75 μl). Clarified mouse plasma free of cells was then subjected to multiplex assays in a Bio-Rad MAGPIX system to define the expression of the noted cytokines in each part before and following treatment (n = 2 studies, eight animals per group ±SEM). *P < 0.05 greater than Day 0 pre-treatment value; **P < 0.05 greater than Day 7 vehicle control; #P < 0.05 less than Day 0 pre-treatment value; ##P < 0.05 less than Day 7 vehicle value; ¶P < 0.05 less than Day 7 regorafenib treatment value. (H) HT29 and HCT116 tumor cells in vitro were treated with vehicle control; [regorafenib (0.5 μM) + FTY720 (50 nM)]; the TGF β receptor inhibitor LY2157299 (0.2, 0.6 μM) or the drugs in combination as indicted for 9 h. After 9 h cell viability was determined in a Hermes WiScan instrument using a live/dead assay (n = 3, ±SEM). *P < 0.05 greater than REGO + FTY value. (I–L) HT29 tumors isolated from vehicle control treated or (regorafenib + sildenafil) treated tumors at the time of animal nadir were subjected to multiplex assays in a Bio-Rad MAGPIX analyzer to determine the expression of cytokines in plasma and the phosphorylation of the indicated signal transduction proteins (n = 8, ±SEM). ~P < 0.05 greater than vehicle control treated tumor value; #P < 0.05 less than vehicle control treated tumor value. (M) HuH7 cells were treated with vehicle control; regorafenib (0.5 μM) and sildenafil (2 μM); BGJ398 (1 μM); Lapatinib (1 μM); MK2206 (1 μM) or in the combinations indicated in the part. Cells were examined 12 h after treatment using a live/dead assay in a Hermes WiScan instrument. Red/yellow cells = dead; green cells = alive (n = 3, ±SEM). *P < 0.05 greater than corresponding value in (regorafenib + sildenafil) treatment alone.
Mentions: Based on our unexpected findings in Figure 9, we determined using a Bio-Rad MAGPIX multiplex system the expression levels of human cytokines in mouse plasma and the expression and activity of signal transduction proteins within the established tumor itself. Regorafenib and sildenafil exposure for 7 days decreased the plasma levels of bFGF and GM-CSF but significantly increased the expression of PDGFbb (Fig. 10A). In agreement with data in Figure 8, treatment of tumors for 7 days with FTY720 increased plasma levels of FAS-L (Fig. 10B). For a number of cytokines, e.g., endoglin, IL6, VEGF-A/C/D, endpoietin 2, EGF, their plasma level declined in vehicle control treated tumors over 7 days of growth which was associated with a mean of approximately sevenfold increase in tumor volume (Fig. 10B–F). In this respect, treatment of tumors with FTY720 or with regorafenib and FTY720 prevented the decline in plasma cytokine levels that were observed in mice with vehicle control treated tumors at day 7 (Fig. 10B–G). For tumors exposed to regorafenib and sildenafil, the expression of pro-growth/pro-angiogenic/pro-invasion cytokines was either unchanged compared to vehicle control treated tumors or was significantly reduced (Fig. 10C–G).

Bottom Line: PDE5 and PDGFRα/β were over-expressed in liver tumors compared to normal liver tissue.Knock down of PDE5 or of PDGFRα/β recapitulated the effects of the individual drugs.The drug combination also reduced mTOR protein expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, Virginia.

Show MeSH
Related in: MedlinePlus