Limits...
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.

Show MeSH

Related in: MedlinePlus

Sorafenib/regorafenib interact in vivo to suppress tumor growth. (A) HuH7 (hepatoma), HCT116 (colorectal) and BT474 (breast) tumors were formed in the flanks of athymic mice (~150 mm3). Animals were treated PO with vehicle diluent (cremophore); sildenafil (5 mg/kg); regorafenib (25 mg/kg); or the drug combination for 24 h. Tumors were isolated and single cell suspensions of tumor cells derived. Cells were plated and colonies permitted to form for 7 days. Colonies were fixed, stained and counted, and the survival value in vehicle treated tumors defined as 100% (n = 3 × 6, ±SEM). #P < 0.05 less than survival in regorafenib treated cells. (B) HuH7 tumors were formed in the flanks of athymic mice (~150 mm3). Animals were treated PO with vehicle diluent (cremophore); sildenafil (5 mg/kg); sorafenib (25 mg/kg); or the drug combination for 3 days. Tumor volumes were measured 7 days and 14 days after the start of drug treatment (n = 2 studies, eight animals per group ±SEM). #P < 0.05 less growth than sorafenib treated cells. (C) HT29 tumors were formed in the flanks of athymic mice (~30 mm3). Animals were 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 for 7 days. Tumor volumes were measured every 4 days after the end of drug treatment (n = 2 studies, eight animals per group ±SEM). (D) Kaplan Meier survival plot of animals treated in part C; animals were humanely sacrificed when tumor volumes exceeded 1,500 mm3. (E) Representative images of tumors from each condition, taken when tumors were of approximately the same volume (between days 15 and 30). (F) HT29 tumors, at the time of sacrifice were isolated from the animals and gently digested to obtain a single cell suspension of tumor cells. Cells were plated as single cells (100–10,000) per well of a 6-well plate. Colonies were permitted to form for 7–10 days after which they were fixed, stained and counted, and the plating efficiency for tumor cells ex vivo for each treatment determined (n = 8, ±SEM). (G) Athymic mice carrying pre-formed HT29 tumors (50 mm3) were treated PO with vehicle diluent (cremophore) or sildenafil (10 mg/kg) and regorafenib (50 mg/kg) for 7 days after which animals were sacrificed, their normal tissues obtained and fixed and sealed in paraffin wax. Ten micron slices of each tissue were taken and H&E stained and examined under 10× magnification.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4835179&req=5

Figure 9: Sorafenib/regorafenib interact in vivo to suppress tumor growth. (A) HuH7 (hepatoma), HCT116 (colorectal) and BT474 (breast) tumors were formed in the flanks of athymic mice (~150 mm3). Animals were treated PO with vehicle diluent (cremophore); sildenafil (5 mg/kg); regorafenib (25 mg/kg); or the drug combination for 24 h. Tumors were isolated and single cell suspensions of tumor cells derived. Cells were plated and colonies permitted to form for 7 days. Colonies were fixed, stained and counted, and the survival value in vehicle treated tumors defined as 100% (n = 3 × 6, ±SEM). #P < 0.05 less than survival in regorafenib treated cells. (B) HuH7 tumors were formed in the flanks of athymic mice (~150 mm3). Animals were treated PO with vehicle diluent (cremophore); sildenafil (5 mg/kg); sorafenib (25 mg/kg); or the drug combination for 3 days. Tumor volumes were measured 7 days and 14 days after the start of drug treatment (n = 2 studies, eight animals per group ±SEM). #P < 0.05 less growth than sorafenib treated cells. (C) HT29 tumors were formed in the flanks of athymic mice (~30 mm3). Animals were 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 for 7 days. Tumor volumes were measured every 4 days after the end of drug treatment (n = 2 studies, eight animals per group ±SEM). (D) Kaplan Meier survival plot of animals treated in part C; animals were humanely sacrificed when tumor volumes exceeded 1,500 mm3. (E) Representative images of tumors from each condition, taken when tumors were of approximately the same volume (between days 15 and 30). (F) HT29 tumors, at the time of sacrifice were isolated from the animals and gently digested to obtain a single cell suspension of tumor cells. Cells were plated as single cells (100–10,000) per well of a 6-well plate. Colonies were permitted to form for 7–10 days after which they were fixed, stained and counted, and the plating efficiency for tumor cells ex vivo for each treatment determined (n = 8, ±SEM). (G) Athymic mice carrying pre-formed HT29 tumors (50 mm3) were treated PO with vehicle diluent (cremophore) or sildenafil (10 mg/kg) and regorafenib (50 mg/kg) for 7 days after which animals were sacrificed, their normal tissues obtained and fixed and sealed in paraffin wax. Ten micron slices of each tissue were taken and H&E stained and examined under 10× magnification.

Mentions: We next determined whether sorafenib/regorafenib interacted with sildenafil in vivo to kill tumor cells. Initial studies examined the ex vivo plating efficiency of tumor cells treated in vivo with regorafenib and sildenafil. Treatment of tumors with regorafenib and sildenafil in vivo caused a reduction in the ex vivo plating efficiency of isolated tumor cells following drug exposure; thus the long-term colony forming ability of drug treated cells was reduced beyond that achieved due to the proximal anti-tumor effects of the drugs (Fig. 9A). Based on the data in Figure 1, we next performed additional studies using pre-formed HuH7 tumors (~150 mm3); in our prior experience HuH7 cells have a significantly greater tumorigenic potential than either HEPG2 or HEP3B cells to grow in an athymic mouse. The a priori prediction for our HuH7 tumor studies would be that due to the lack of CD95 expression, the relative amount of killing may be more modest, although data from Figure 9A argues that in vivo killing by our regorafenib and sildenafil combination occurs. Transient treatment of animals with sildenafil did not significantly alter tumor growth whereas transient treatment with sorafenib caused a non-significant trend towards reduced growth (Fig. 9B). Combined treatment of animals with both drugs for 3 days caused a significant reduction/delay in HuH7 tumor re-growth.


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)

Sorafenib/regorafenib interact in vivo to suppress tumor growth. (A) HuH7 (hepatoma), HCT116 (colorectal) and BT474 (breast) tumors were formed in the flanks of athymic mice (~150 mm3). Animals were treated PO with vehicle diluent (cremophore); sildenafil (5 mg/kg); regorafenib (25 mg/kg); or the drug combination for 24 h. Tumors were isolated and single cell suspensions of tumor cells derived. Cells were plated and colonies permitted to form for 7 days. Colonies were fixed, stained and counted, and the survival value in vehicle treated tumors defined as 100% (n = 3 × 6, ±SEM). #P < 0.05 less than survival in regorafenib treated cells. (B) HuH7 tumors were formed in the flanks of athymic mice (~150 mm3). Animals were treated PO with vehicle diluent (cremophore); sildenafil (5 mg/kg); sorafenib (25 mg/kg); or the drug combination for 3 days. Tumor volumes were measured 7 days and 14 days after the start of drug treatment (n = 2 studies, eight animals per group ±SEM). #P < 0.05 less growth than sorafenib treated cells. (C) HT29 tumors were formed in the flanks of athymic mice (~30 mm3). Animals were 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 for 7 days. Tumor volumes were measured every 4 days after the end of drug treatment (n = 2 studies, eight animals per group ±SEM). (D) Kaplan Meier survival plot of animals treated in part C; animals were humanely sacrificed when tumor volumes exceeded 1,500 mm3. (E) Representative images of tumors from each condition, taken when tumors were of approximately the same volume (between days 15 and 30). (F) HT29 tumors, at the time of sacrifice were isolated from the animals and gently digested to obtain a single cell suspension of tumor cells. Cells were plated as single cells (100–10,000) per well of a 6-well plate. Colonies were permitted to form for 7–10 days after which they were fixed, stained and counted, and the plating efficiency for tumor cells ex vivo for each treatment determined (n = 8, ±SEM). (G) Athymic mice carrying pre-formed HT29 tumors (50 mm3) were treated PO with vehicle diluent (cremophore) or sildenafil (10 mg/kg) and regorafenib (50 mg/kg) for 7 days after which animals were sacrificed, their normal tissues obtained and fixed and sealed in paraffin wax. Ten micron slices of each tissue were taken and H&E stained and examined under 10× magnification.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4835179&req=5

Figure 9: Sorafenib/regorafenib interact in vivo to suppress tumor growth. (A) HuH7 (hepatoma), HCT116 (colorectal) and BT474 (breast) tumors were formed in the flanks of athymic mice (~150 mm3). Animals were treated PO with vehicle diluent (cremophore); sildenafil (5 mg/kg); regorafenib (25 mg/kg); or the drug combination for 24 h. Tumors were isolated and single cell suspensions of tumor cells derived. Cells were plated and colonies permitted to form for 7 days. Colonies were fixed, stained and counted, and the survival value in vehicle treated tumors defined as 100% (n = 3 × 6, ±SEM). #P < 0.05 less than survival in regorafenib treated cells. (B) HuH7 tumors were formed in the flanks of athymic mice (~150 mm3). Animals were treated PO with vehicle diluent (cremophore); sildenafil (5 mg/kg); sorafenib (25 mg/kg); or the drug combination for 3 days. Tumor volumes were measured 7 days and 14 days after the start of drug treatment (n = 2 studies, eight animals per group ±SEM). #P < 0.05 less growth than sorafenib treated cells. (C) HT29 tumors were formed in the flanks of athymic mice (~30 mm3). Animals were 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 for 7 days. Tumor volumes were measured every 4 days after the end of drug treatment (n = 2 studies, eight animals per group ±SEM). (D) Kaplan Meier survival plot of animals treated in part C; animals were humanely sacrificed when tumor volumes exceeded 1,500 mm3. (E) Representative images of tumors from each condition, taken when tumors were of approximately the same volume (between days 15 and 30). (F) HT29 tumors, at the time of sacrifice were isolated from the animals and gently digested to obtain a single cell suspension of tumor cells. Cells were plated as single cells (100–10,000) per well of a 6-well plate. Colonies were permitted to form for 7–10 days after which they were fixed, stained and counted, and the plating efficiency for tumor cells ex vivo for each treatment determined (n = 8, ±SEM). (G) Athymic mice carrying pre-formed HT29 tumors (50 mm3) were treated PO with vehicle diluent (cremophore) or sildenafil (10 mg/kg) and regorafenib (50 mg/kg) for 7 days after which animals were sacrificed, their normal tissues obtained and fixed and sealed in paraffin wax. Ten micron slices of each tissue were taken and H&E stained and examined under 10× magnification.
Mentions: We next determined whether sorafenib/regorafenib interacted with sildenafil in vivo to kill tumor cells. Initial studies examined the ex vivo plating efficiency of tumor cells treated in vivo with regorafenib and sildenafil. Treatment of tumors with regorafenib and sildenafil in vivo caused a reduction in the ex vivo plating efficiency of isolated tumor cells following drug exposure; thus the long-term colony forming ability of drug treated cells was reduced beyond that achieved due to the proximal anti-tumor effects of the drugs (Fig. 9A). Based on the data in Figure 1, we next performed additional studies using pre-formed HuH7 tumors (~150 mm3); in our prior experience HuH7 cells have a significantly greater tumorigenic potential than either HEPG2 or HEP3B cells to grow in an athymic mouse. The a priori prediction for our HuH7 tumor studies would be that due to the lack of CD95 expression, the relative amount of killing may be more modest, although data from Figure 9A argues that in vivo killing by our regorafenib and sildenafil combination occurs. Transient treatment of animals with sildenafil did not significantly alter tumor growth whereas transient treatment with sorafenib caused a non-significant trend towards reduced growth (Fig. 9B). Combined treatment of animals with both drugs for 3 days caused a significant reduction/delay in HuH7 tumor re-growth.

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