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OSU-03012 and Viagra Treatment Inhibits the Activity of Multiple Chaperone Proteins and Disrupts the Blood-Brain Barrier: Implications for Anti-Cancer Therapies.

Booth L, Roberts JL, Tavallai M, Nourbakhsh A, Chuckalovcak J, Carter J, Poklepovic A, Dent P - J. Cell. Physiol. (2015)

Bottom Line: Sildenafil (Viagra) interacted in a greater than additive fashion with OSU-03012 to kill stem-like GBM cells.In multiplex assays on plasma and human tumor tissue from an OSU-03012/sildenafil treated mouse, we noted a profound reduction in uPA signaling and identified FGF and JAK1/2 as response biomarkers for potentially suppressing the killing response.Inhibition of FGFR signaling and to a lesser extent JAK1/2 signaling profoundly enhanced OSU-03012/sildenafil lethality.

View Article: PubMed Central - PubMed

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

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Treatment of tumor cells with [OSU-03012 + sildenafil] or [celecoxib + sildenafil] rapidly reduces the expression of multiple growth factor receptors. A: BT474 mammary carcinoma cells known to strongly over-express ERBB2 and: B–F: GBM5/6/12/14 cells; in 96-well plates were treated with vehicle; celecoxib (5 μM) + sildenafil (2 μM); or OSU-03012 (1 μM) + sildenafil and after 6 h treatment were fixed with 2% (v/v) paraformaldehyde containing Triton X100 to permeabilized cells. Immunostaining of the indicated growth factor receptors and plasma membrane drug transporters was performed using standard techniques and IF images detected using a Hermes Wiscan machine. G,H: Normal tissues from mice treated in Figure 1section (D), brain and liver, were fixed, sectioned and immune-stained to determine the total expression of GRP78, ABCB1, and ABCG2. Images were examined under 10× magnification using a Hermes WiScan instrument.
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Figure 2: Treatment of tumor cells with [OSU-03012 + sildenafil] or [celecoxib + sildenafil] rapidly reduces the expression of multiple growth factor receptors. A: BT474 mammary carcinoma cells known to strongly over-express ERBB2 and: B–F: GBM5/6/12/14 cells; in 96-well plates were treated with vehicle; celecoxib (5 μM) + sildenafil (2 μM); or OSU-03012 (1 μM) + sildenafil and after 6 h treatment were fixed with 2% (v/v) paraformaldehyde containing Triton X100 to permeabilized cells. Immunostaining of the indicated growth factor receptors and plasma membrane drug transporters was performed using standard techniques and IF images detected using a Hermes Wiscan machine. G,H: Normal tissues from mice treated in Figure 1section (D), brain and liver, were fixed, sectioned and immune-stained to determine the total expression of GRP78, ABCB1, and ABCG2. Images were examined under 10× magnification using a Hermes WiScan instrument.

Mentions: Expression of ERBB1, ERBB2, and of ERBB3 and ERBB4 in BT474 breast cancer cells was reduced by OSU + SIL treatment (Fig. 2A). Treatment of GBM cells with CEL + SIL and with OSU + SIL rapidly reduced total expression of ERBB1 (GBM12) and ERBB1 vIII (GBM6) within 6 h, whereas expression of c-MET was reduced to a much lesser extent and in a cell-dependent fashion (Fig. 2B). Drug combination exposures rapidly reduced expression of S1PR1 (EDG1) and S1PR5 (EDG5) as they did in a cell type dependent fashion for the IL6R and the IL8R (Fig. 2C,D). Expression of the IGF1R and of TNFαR1 were also reduced by the drug combination (Fig. 2E). Drug combination treatment also reduced the total expression of multiple plasma membrane associated drug efflux pumps; pumps that are generally over-expressed in recurrent patient tumors and are partly responsible for chemotherapy resistance as well as for a functional blood–brain barrier (Fig. 2F). Identical data were obtained in vivo examining normal brain and liver tissues (Fig. 2G,H). Similar data showing knock down of ABCB1 and ABCG2 expression were also obtained in stem cell selected variants of GBM5/6/12/14 (data not shown). In previous studies, we have shown that expression of the death receptor CD95 (FAS) does not alter after particular these drug exposures and in fact becomes plasma membrane localized and clustered, observations indicative of CD95 activation (Booth et al., 2014a,b).


OSU-03012 and Viagra Treatment Inhibits the Activity of Multiple Chaperone Proteins and Disrupts the Blood-Brain Barrier: Implications for Anti-Cancer Therapies.

Booth L, Roberts JL, Tavallai M, Nourbakhsh A, Chuckalovcak J, Carter J, Poklepovic A, Dent P - J. Cell. Physiol. (2015)

Treatment of tumor cells with [OSU-03012 + sildenafil] or [celecoxib + sildenafil] rapidly reduces the expression of multiple growth factor receptors. A: BT474 mammary carcinoma cells known to strongly over-express ERBB2 and: B–F: GBM5/6/12/14 cells; in 96-well plates were treated with vehicle; celecoxib (5 μM) + sildenafil (2 μM); or OSU-03012 (1 μM) + sildenafil and after 6 h treatment were fixed with 2% (v/v) paraformaldehyde containing Triton X100 to permeabilized cells. Immunostaining of the indicated growth factor receptors and plasma membrane drug transporters was performed using standard techniques and IF images detected using a Hermes Wiscan machine. G,H: Normal tissues from mice treated in Figure 1section (D), brain and liver, were fixed, sectioned and immune-stained to determine the total expression of GRP78, ABCB1, and ABCG2. Images were examined under 10× magnification using a Hermes WiScan instrument.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4835175&req=5

Figure 2: Treatment of tumor cells with [OSU-03012 + sildenafil] or [celecoxib + sildenafil] rapidly reduces the expression of multiple growth factor receptors. A: BT474 mammary carcinoma cells known to strongly over-express ERBB2 and: B–F: GBM5/6/12/14 cells; in 96-well plates were treated with vehicle; celecoxib (5 μM) + sildenafil (2 μM); or OSU-03012 (1 μM) + sildenafil and after 6 h treatment were fixed with 2% (v/v) paraformaldehyde containing Triton X100 to permeabilized cells. Immunostaining of the indicated growth factor receptors and plasma membrane drug transporters was performed using standard techniques and IF images detected using a Hermes Wiscan machine. G,H: Normal tissues from mice treated in Figure 1section (D), brain and liver, were fixed, sectioned and immune-stained to determine the total expression of GRP78, ABCB1, and ABCG2. Images were examined under 10× magnification using a Hermes WiScan instrument.
Mentions: Expression of ERBB1, ERBB2, and of ERBB3 and ERBB4 in BT474 breast cancer cells was reduced by OSU + SIL treatment (Fig. 2A). Treatment of GBM cells with CEL + SIL and with OSU + SIL rapidly reduced total expression of ERBB1 (GBM12) and ERBB1 vIII (GBM6) within 6 h, whereas expression of c-MET was reduced to a much lesser extent and in a cell-dependent fashion (Fig. 2B). Drug combination exposures rapidly reduced expression of S1PR1 (EDG1) and S1PR5 (EDG5) as they did in a cell type dependent fashion for the IL6R and the IL8R (Fig. 2C,D). Expression of the IGF1R and of TNFαR1 were also reduced by the drug combination (Fig. 2E). Drug combination treatment also reduced the total expression of multiple plasma membrane associated drug efflux pumps; pumps that are generally over-expressed in recurrent patient tumors and are partly responsible for chemotherapy resistance as well as for a functional blood–brain barrier (Fig. 2F). Identical data were obtained in vivo examining normal brain and liver tissues (Fig. 2G,H). Similar data showing knock down of ABCB1 and ABCG2 expression were also obtained in stem cell selected variants of GBM5/6/12/14 (data not shown). In previous studies, we have shown that expression of the death receptor CD95 (FAS) does not alter after particular these drug exposures and in fact becomes plasma membrane localized and clustered, observations indicative of CD95 activation (Booth et al., 2014a,b).

Bottom Line: Sildenafil (Viagra) interacted in a greater than additive fashion with OSU-03012 to kill stem-like GBM cells.In multiplex assays on plasma and human tumor tissue from an OSU-03012/sildenafil treated mouse, we noted a profound reduction in uPA signaling and identified FGF and JAK1/2 as response biomarkers for potentially suppressing the killing response.Inhibition of FGFR signaling and to a lesser extent JAK1/2 signaling profoundly enhanced OSU-03012/sildenafil lethality.

View Article: PubMed Central - PubMed

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

Show MeSH
Related in: MedlinePlus