Limits...
Sulindac derivatives inhibit cell growth and induce apoptosis in primary cells from malignant peripheral nerve sheath tumors of NF1-patients.

Frahm S, Kurtz A, Kluwe L, Farassati F, Friedrich RE, Mautner VF - Cancer Cell Int. (2004)

Bottom Line: The decrease in viability of the tested cells correlated with induction of apoptosis.Treatment with 500 microM Exisulind and 125 microM Sulindac Sulfide for a period of 2 days increased the rate of apoptosis 21-27-fold compared to untreated cells.Moreover, elevated levels of phosphorylated SAPK/JNK were found after drug treatment, and low activation of cleaved caspase-3 was seen.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Brain Tumor Biology, Department of Neurosurgery, University Hospital Eppendorf, Hamburg, Germany. sfrahm@uke.uni-hamburg.de

ABSTRACT
BACKGROUND: Malignant peripheral nerve sheath tumors (MPNSTs) are neoplasms leading to death in most cases. Patients with Neurofibromatosis type 1 have an increased risk of developing this malignancy. The metabolites of the inactive prodrug Sulindac, Sulindac Sulfide and Sulindac Sulfone (Exisulind) are new chemopreventive agents that show promising results in the treatment of different cancer types. In this study we examined the antineoplastic effect of these compounds on primary cells derived from two MPNSTs of Neurofibromatosis type 1 patients. RESULTS: Exisulind and Sulindac Sulfide showed a dramatic time- and dose-dependent growth inhibitory effect with IC50-values of 120 microM and 63 microM, respectively. The decrease in viability of the tested cells correlated with induction of apoptosis. Treatment with 500 microM Exisulind and 125 microM Sulindac Sulfide for a period of 2 days increased the rate of apoptosis 21-27-fold compared to untreated cells. Reduced expression of RAS-GTP and phosphorylated ERK1/2 was detected in treated MPNST cells. Moreover, elevated levels of phosphorylated SAPK/JNK were found after drug treatment, and low activation of cleaved caspase-3 was seen. CONCLUSIONS: Our results suggest that this class of compounds may be of therapeutic benefit for Neurofibromatosis type 1 patients with MPNST.

No MeSH data available.


Related in: MedlinePlus

Inhibition of RAS-GTP and phosphorylated ERK1/2 in human MPNST cells exposed to Sulindac derivatives A: Cell lines S462 and S520 were grown in DMEM with 10% serum and treated at 80–90% confluency with either 0.2% DMSO, 500 μM Exisulind (Exi) or 125 μM Sulindac Sulfide (s. s.). Cells were lysed after 24 h and RAS was immunoprecipitated and detected by western blotting with an anti-RAS antibody, the same lysates were blotted for phosphorylated (phospho-) and basal (total-) ERK1/2. B: Upregulation of phosphorylated ERK1/2 and AKT for treated and untreated cells after addition of EGF, whereas the RAS-GTP level remains unchanged at all conditions. Cell line S462 was starved over night and then Sulindac metabolites were supplemented at concentrations mentioned above in DMEM containing 0.1% serum. EGF was added after 24 h of treatment, cells were lysed 15 min later and after western blotting incubated with phosphorylated and unphosphorylated ERK1/2, phospho-AKT and RAS-antibodies.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC425591&req=5

Figure 5: Inhibition of RAS-GTP and phosphorylated ERK1/2 in human MPNST cells exposed to Sulindac derivatives A: Cell lines S462 and S520 were grown in DMEM with 10% serum and treated at 80–90% confluency with either 0.2% DMSO, 500 μM Exisulind (Exi) or 125 μM Sulindac Sulfide (s. s.). Cells were lysed after 24 h and RAS was immunoprecipitated and detected by western blotting with an anti-RAS antibody, the same lysates were blotted for phosphorylated (phospho-) and basal (total-) ERK1/2. B: Upregulation of phosphorylated ERK1/2 and AKT for treated and untreated cells after addition of EGF, whereas the RAS-GTP level remains unchanged at all conditions. Cell line S462 was starved over night and then Sulindac metabolites were supplemented at concentrations mentioned above in DMEM containing 0.1% serum. EGF was added after 24 h of treatment, cells were lysed 15 min later and after western blotting incubated with phosphorylated and unphosphorylated ERK1/2, phospho-AKT and RAS-antibodies.

Mentions: To ascertain the effect of the Sulindac metabolites on the RAS signaling pathway, we determined the levels of RAS-GTP by affinity precipitation and western blotting with a monoclonal RAS antibody, as well as activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2). After 24 h treatment with the Sulindac metabolites, reduction of RAS-GTP levels were observed in both cell lines, slightly more pronounced in Exisulind treated cells (Fig. 5a). In concordance, phosphorylation of ERK1 (p44) and ERK2 (p42) was also reduced after the treatment. No changes in basal ERK1/2 levels were observed (Fig. 5a). Under serum-free conditions a significant reduction of phospho-ERK1/2 activation was detected already after 8 h of treatment (data not shown).


Sulindac derivatives inhibit cell growth and induce apoptosis in primary cells from malignant peripheral nerve sheath tumors of NF1-patients.

Frahm S, Kurtz A, Kluwe L, Farassati F, Friedrich RE, Mautner VF - Cancer Cell Int. (2004)

Inhibition of RAS-GTP and phosphorylated ERK1/2 in human MPNST cells exposed to Sulindac derivatives A: Cell lines S462 and S520 were grown in DMEM with 10% serum and treated at 80–90% confluency with either 0.2% DMSO, 500 μM Exisulind (Exi) or 125 μM Sulindac Sulfide (s. s.). Cells were lysed after 24 h and RAS was immunoprecipitated and detected by western blotting with an anti-RAS antibody, the same lysates were blotted for phosphorylated (phospho-) and basal (total-) ERK1/2. B: Upregulation of phosphorylated ERK1/2 and AKT for treated and untreated cells after addition of EGF, whereas the RAS-GTP level remains unchanged at all conditions. Cell line S462 was starved over night and then Sulindac metabolites were supplemented at concentrations mentioned above in DMEM containing 0.1% serum. EGF was added after 24 h of treatment, cells were lysed 15 min later and after western blotting incubated with phosphorylated and unphosphorylated ERK1/2, phospho-AKT and RAS-antibodies.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC425591&req=5

Figure 5: Inhibition of RAS-GTP and phosphorylated ERK1/2 in human MPNST cells exposed to Sulindac derivatives A: Cell lines S462 and S520 were grown in DMEM with 10% serum and treated at 80–90% confluency with either 0.2% DMSO, 500 μM Exisulind (Exi) or 125 μM Sulindac Sulfide (s. s.). Cells were lysed after 24 h and RAS was immunoprecipitated and detected by western blotting with an anti-RAS antibody, the same lysates were blotted for phosphorylated (phospho-) and basal (total-) ERK1/2. B: Upregulation of phosphorylated ERK1/2 and AKT for treated and untreated cells after addition of EGF, whereas the RAS-GTP level remains unchanged at all conditions. Cell line S462 was starved over night and then Sulindac metabolites were supplemented at concentrations mentioned above in DMEM containing 0.1% serum. EGF was added after 24 h of treatment, cells were lysed 15 min later and after western blotting incubated with phosphorylated and unphosphorylated ERK1/2, phospho-AKT and RAS-antibodies.
Mentions: To ascertain the effect of the Sulindac metabolites on the RAS signaling pathway, we determined the levels of RAS-GTP by affinity precipitation and western blotting with a monoclonal RAS antibody, as well as activation of extracellular signal-regulated kinases 1 and 2 (ERK1/2). After 24 h treatment with the Sulindac metabolites, reduction of RAS-GTP levels were observed in both cell lines, slightly more pronounced in Exisulind treated cells (Fig. 5a). In concordance, phosphorylation of ERK1 (p44) and ERK2 (p42) was also reduced after the treatment. No changes in basal ERK1/2 levels were observed (Fig. 5a). Under serum-free conditions a significant reduction of phospho-ERK1/2 activation was detected already after 8 h of treatment (data not shown).

Bottom Line: The decrease in viability of the tested cells correlated with induction of apoptosis.Treatment with 500 microM Exisulind and 125 microM Sulindac Sulfide for a period of 2 days increased the rate of apoptosis 21-27-fold compared to untreated cells.Moreover, elevated levels of phosphorylated SAPK/JNK were found after drug treatment, and low activation of cleaved caspase-3 was seen.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Brain Tumor Biology, Department of Neurosurgery, University Hospital Eppendorf, Hamburg, Germany. sfrahm@uke.uni-hamburg.de

ABSTRACT
BACKGROUND: Malignant peripheral nerve sheath tumors (MPNSTs) are neoplasms leading to death in most cases. Patients with Neurofibromatosis type 1 have an increased risk of developing this malignancy. The metabolites of the inactive prodrug Sulindac, Sulindac Sulfide and Sulindac Sulfone (Exisulind) are new chemopreventive agents that show promising results in the treatment of different cancer types. In this study we examined the antineoplastic effect of these compounds on primary cells derived from two MPNSTs of Neurofibromatosis type 1 patients. RESULTS: Exisulind and Sulindac Sulfide showed a dramatic time- and dose-dependent growth inhibitory effect with IC50-values of 120 microM and 63 microM, respectively. The decrease in viability of the tested cells correlated with induction of apoptosis. Treatment with 500 microM Exisulind and 125 microM Sulindac Sulfide for a period of 2 days increased the rate of apoptosis 21-27-fold compared to untreated cells. Reduced expression of RAS-GTP and phosphorylated ERK1/2 was detected in treated MPNST cells. Moreover, elevated levels of phosphorylated SAPK/JNK were found after drug treatment, and low activation of cleaved caspase-3 was seen. CONCLUSIONS: Our results suggest that this class of compounds may be of therapeutic benefit for Neurofibromatosis type 1 patients with MPNST.

No MeSH data available.


Related in: MedlinePlus