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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

ROS/RNS regulate the induction of autophagy by drug treatment. (A and B) HEPG2 cells were transfected with a plasmid to express LC3-GFP-RFP. Twenty-four hours after transfection cells were, as indicated pre-treated with L-NAME (1 μM) or NAC (10 mM), then treated with vehicle (DMSO), sorafenib (2 μM), sildenafil (2 μM); or the drugs in combination. The number of GFP vesicles (early autophagosomes) and RFP vesicles (late autolysosomes) were determined 6 and 24 h after drug treatment (n = 3, ±SEM). #P < 0.05 less than corresponding value in cells not treated with L-NAME or NAC. (C) HEP3B and HuH7 cells were transfected with a control scrambled siRNA (siSCR) or siRNA molecules to knock down expression of Beclin1 or ATG5. Thirty-six hours after transfection were treated with vehicle or with sildenafil (2 μM), and/or sorafenib (2 μM). Twenty-four hours after treatment cells were isolated and viability determined by trypan blue (n = 3, ±SEM). #P < 0.05 less than corresponding value in siSCR cells. (D) Lower graph: HEPG2 cells were transfected with a control scrambled siRNA (siSCR) or siRNA molecules to knock down expression of Beclin1, ULK1, or ATG5. Thirty-six hours after transfection were treated with vehicle or with sildenafil (2 μM), and/or regorafenib (0.5 μM). Twenty-four hours after treatment cells were isolated and viability determine by trypan blue (n = 3, ±SEM); Middle images: Cells expressing LC3-GFP were treated with vehicle or with sildenafil (2 μM), and/or regorafenib (0.5 μM) and the number of intense GFP+ vesicles counted in 40 cells and the mean number of vesicles per cell is presented (n = 3, ±SEM); Upper blots: Cells were treated with vehicle or with sildenafil (2 μM), and/or regorafenib (0.5 μM) and isolated after 6 h. Cell lysates were western blotted for the expression of the indicated proteins (n = 3).
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Figure 5: ROS/RNS regulate the induction of autophagy by drug treatment. (A and B) HEPG2 cells were transfected with a plasmid to express LC3-GFP-RFP. Twenty-four hours after transfection cells were, as indicated pre-treated with L-NAME (1 μM) or NAC (10 mM), then treated with vehicle (DMSO), sorafenib (2 μM), sildenafil (2 μM); or the drugs in combination. The number of GFP vesicles (early autophagosomes) and RFP vesicles (late autolysosomes) were determined 6 and 24 h after drug treatment (n = 3, ±SEM). #P < 0.05 less than corresponding value in cells not treated with L-NAME or NAC. (C) HEP3B and HuH7 cells were transfected with a control scrambled siRNA (siSCR) or siRNA molecules to knock down expression of Beclin1 or ATG5. Thirty-six hours after transfection were treated with vehicle or with sildenafil (2 μM), and/or sorafenib (2 μM). Twenty-four hours after treatment cells were isolated and viability determined by trypan blue (n = 3, ±SEM). #P < 0.05 less than corresponding value in siSCR cells. (D) Lower graph: HEPG2 cells were transfected with a control scrambled siRNA (siSCR) or siRNA molecules to knock down expression of Beclin1, ULK1, or ATG5. Thirty-six hours after transfection were treated with vehicle or with sildenafil (2 μM), and/or regorafenib (0.5 μM). Twenty-four hours after treatment cells were isolated and viability determine by trypan blue (n = 3, ±SEM); Middle images: Cells expressing LC3-GFP were treated with vehicle or with sildenafil (2 μM), and/or regorafenib (0.5 μM) and the number of intense GFP+ vesicles counted in 40 cells and the mean number of vesicles per cell is presented (n = 3, ±SEM); Upper blots: Cells were treated with vehicle or with sildenafil (2 μM), and/or regorafenib (0.5 μM) and isolated after 6 h. Cell lysates were western blotted for the expression of the indicated proteins (n = 3).

Mentions: In prior studies, we have shown that sorafenib as a single agent can modestly promote increased numbers of autophagosomes (Park et al., 2008, 2010a,b; Martin et al., 2009; Cruickshanks et al., 2013). Treatment of cells with sildenafil and sorafenib increased the numbers of autophagosomes and autolysosomes in a greater than additive fashion (Fig. 5A and B). The increase in the numbers of autophagosomes and autolysosomes was suppressed by incubation of cells with either L-NAME or NAC. Knock down of either ULK-1, Beclin1, or ATG5 expression inhibited the drug-induced increase in autophagosome and autolysosome levels and reduced cell killing by sildenafil and sorafenib or by sildenafil and regorafenib treatments (Fig. 5C and D).


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)

ROS/RNS regulate the induction of autophagy by drug treatment. (A and B) HEPG2 cells were transfected with a plasmid to express LC3-GFP-RFP. Twenty-four hours after transfection cells were, as indicated pre-treated with L-NAME (1 μM) or NAC (10 mM), then treated with vehicle (DMSO), sorafenib (2 μM), sildenafil (2 μM); or the drugs in combination. The number of GFP vesicles (early autophagosomes) and RFP vesicles (late autolysosomes) were determined 6 and 24 h after drug treatment (n = 3, ±SEM). #P < 0.05 less than corresponding value in cells not treated with L-NAME or NAC. (C) HEP3B and HuH7 cells were transfected with a control scrambled siRNA (siSCR) or siRNA molecules to knock down expression of Beclin1 or ATG5. Thirty-six hours after transfection were treated with vehicle or with sildenafil (2 μM), and/or sorafenib (2 μM). Twenty-four hours after treatment cells were isolated and viability determined by trypan blue (n = 3, ±SEM). #P < 0.05 less than corresponding value in siSCR cells. (D) Lower graph: HEPG2 cells were transfected with a control scrambled siRNA (siSCR) or siRNA molecules to knock down expression of Beclin1, ULK1, or ATG5. Thirty-six hours after transfection were treated with vehicle or with sildenafil (2 μM), and/or regorafenib (0.5 μM). Twenty-four hours after treatment cells were isolated and viability determine by trypan blue (n = 3, ±SEM); Middle images: Cells expressing LC3-GFP were treated with vehicle or with sildenafil (2 μM), and/or regorafenib (0.5 μM) and the number of intense GFP+ vesicles counted in 40 cells and the mean number of vesicles per cell is presented (n = 3, ±SEM); Upper blots: Cells were treated with vehicle or with sildenafil (2 μM), and/or regorafenib (0.5 μM) and isolated after 6 h. Cell lysates were western blotted for the expression of the indicated proteins (n = 3).
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Related In: Results  -  Collection

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Figure 5: ROS/RNS regulate the induction of autophagy by drug treatment. (A and B) HEPG2 cells were transfected with a plasmid to express LC3-GFP-RFP. Twenty-four hours after transfection cells were, as indicated pre-treated with L-NAME (1 μM) or NAC (10 mM), then treated with vehicle (DMSO), sorafenib (2 μM), sildenafil (2 μM); or the drugs in combination. The number of GFP vesicles (early autophagosomes) and RFP vesicles (late autolysosomes) were determined 6 and 24 h after drug treatment (n = 3, ±SEM). #P < 0.05 less than corresponding value in cells not treated with L-NAME or NAC. (C) HEP3B and HuH7 cells were transfected with a control scrambled siRNA (siSCR) or siRNA molecules to knock down expression of Beclin1 or ATG5. Thirty-six hours after transfection were treated with vehicle or with sildenafil (2 μM), and/or sorafenib (2 μM). Twenty-four hours after treatment cells were isolated and viability determined by trypan blue (n = 3, ±SEM). #P < 0.05 less than corresponding value in siSCR cells. (D) Lower graph: HEPG2 cells were transfected with a control scrambled siRNA (siSCR) or siRNA molecules to knock down expression of Beclin1, ULK1, or ATG5. Thirty-six hours after transfection were treated with vehicle or with sildenafil (2 μM), and/or regorafenib (0.5 μM). Twenty-four hours after treatment cells were isolated and viability determine by trypan blue (n = 3, ±SEM); Middle images: Cells expressing LC3-GFP were treated with vehicle or with sildenafil (2 μM), and/or regorafenib (0.5 μM) and the number of intense GFP+ vesicles counted in 40 cells and the mean number of vesicles per cell is presented (n = 3, ±SEM); Upper blots: Cells were treated with vehicle or with sildenafil (2 μM), and/or regorafenib (0.5 μM) and isolated after 6 h. Cell lysates were western blotted for the expression of the indicated proteins (n = 3).
Mentions: In prior studies, we have shown that sorafenib as a single agent can modestly promote increased numbers of autophagosomes (Park et al., 2008, 2010a,b; Martin et al., 2009; Cruickshanks et al., 2013). Treatment of cells with sildenafil and sorafenib increased the numbers of autophagosomes and autolysosomes in a greater than additive fashion (Fig. 5A and B). The increase in the numbers of autophagosomes and autolysosomes was suppressed by incubation of cells with either L-NAME or NAC. Knock down of either ULK-1, Beclin1, or ATG5 expression inhibited the drug-induced increase in autophagosome and autolysosome levels and reduced cell killing by sildenafil and sorafenib or by sildenafil and regorafenib treatments (Fig. 5C and D).

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