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Juglone induces apoptosis of tumor stem-like cells through ROS-p38 pathway in glioblastoma

View Article: PubMed Central - PubMed

ABSTRACT

Background: Juglone is a natural pigment, which has cytotoxic effect against various human tumor cells. However, its cytotoxicity to glioma cells, especially to tumor stem-like cells (TSCs) has not been demonstrated.

Methods: TSCs of glioma were enriched from U87 and two primary cells (SHG62, and SHG66) using serum-free medium supplemented with growth factors, including bFGF, EGF and B27. After treatment of juglone with gradient concentrations (0, 10, 20, and 40 μM), the viability and apoptosis of TSCs were evaluated by WST-8 assay and flow cytometry. Reactive oxygen species (ROS) was labeled by the cell-permeable fluorescent probe and detected with flow cytometry. ROS scavenger (NAC) and p38-MAPK inhibitor (SB203580) were applied to resist the cytotoxic effect. Caspase 9 cleavage and p38 phosphorylation (P-p38) were quantified by western blot. Juglone as well as temozolomide (TMZ) were administrated in intracranial xenografts and MR scan was performed every week to evaluate the anti-tumor effect in vivo.

Results: Juglone could obviously inhibit the proliferation of TSCs in glioma by decreasing cell viability (P < 0.01) and inducing apoptosis (P < 0.01), which was accompanied by increased caspase 9 cleavage in a dose-dependent manner (P < 0.01). In the meantime, juglone could generate ROS significantly and increase p38 phosphorylation (P < 0.01). In addition, pretreatment with ROS scavenger or p38-MAPK inhibitor could reverse juglone-induced cytotoxicity (P < 0.01). More importantly, juglone could also suppress tumor growth in vivo and improve the survival of U87-bearing mice compared with control (P < 0.05), although TMZ seemed to have better effect.

Conclusions: Juglone could inhibit the growth of TSCs in gliomas through the activation of ROS-p38-MAPK pathway in vitro, and the anti-glioma effect was validated in vivo, which offers a potential therapeutic agent to gliomas.

No MeSH data available.


Juglone could generate ROS and activate p38 phosphorylation. a Flow cytometry showed that juglone-induced ROS generation was increased in a dose-dependent manner. b Statistical data of ROS MFI in different groups (**P < 0.01). c Juglone treatment significantly increased p38 phosphorylation in a dose-dependent manner. d Statistical data of P-p38 protein at different concentrations using western blot (**P < 0.01)
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Fig3: Juglone could generate ROS and activate p38 phosphorylation. a Flow cytometry showed that juglone-induced ROS generation was increased in a dose-dependent manner. b Statistical data of ROS MFI in different groups (**P < 0.01). c Juglone treatment significantly increased p38 phosphorylation in a dose-dependent manner. d Statistical data of P-p38 protein at different concentrations using western blot (**P < 0.01)

Mentions: Juglone-induced ROS generation was obvious in a dose-dependent manneras compared to vehicle control (P < 0.01) (Fig. 3a and b). As demonstrated by western blot (Fig. 3c and d), juglone (20, 40 μM) treatment significantly increased p38 phosphorylation (P < 0.01),which indicated p38-MAPK pathway activation in TSCs.Fig. 3


Juglone induces apoptosis of tumor stem-like cells through ROS-p38 pathway in glioblastoma
Juglone could generate ROS and activate p38 phosphorylation. a Flow cytometry showed that juglone-induced ROS generation was increased in a dose-dependent manner. b Statistical data of ROS MFI in different groups (**P < 0.01). c Juglone treatment significantly increased p38 phosphorylation in a dose-dependent manner. d Statistical data of P-p38 protein at different concentrations using western blot (**P < 0.01)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC5383964&req=5

Fig3: Juglone could generate ROS and activate p38 phosphorylation. a Flow cytometry showed that juglone-induced ROS generation was increased in a dose-dependent manner. b Statistical data of ROS MFI in different groups (**P < 0.01). c Juglone treatment significantly increased p38 phosphorylation in a dose-dependent manner. d Statistical data of P-p38 protein at different concentrations using western blot (**P < 0.01)
Mentions: Juglone-induced ROS generation was obvious in a dose-dependent manneras compared to vehicle control (P < 0.01) (Fig. 3a and b). As demonstrated by western blot (Fig. 3c and d), juglone (20, 40 μM) treatment significantly increased p38 phosphorylation (P < 0.01),which indicated p38-MAPK pathway activation in TSCs.Fig. 3

View Article: PubMed Central - PubMed

ABSTRACT

Background: Juglone is a natural pigment, which has cytotoxic effect against various human tumor cells. However, its cytotoxicity to glioma cells, especially to tumor stem-like cells (TSCs) has not been demonstrated.

Methods: TSCs of glioma were enriched from U87 and two primary cells (SHG62, and SHG66) using serum-free medium supplemented with growth factors, including bFGF, EGF and B27. After treatment of juglone with gradient concentrations (0, 10, 20, and 40&nbsp;&mu;M), the viability and apoptosis of TSCs were evaluated by WST-8 assay and flow cytometry. Reactive oxygen species (ROS) was labeled by the cell-permeable fluorescent probe and detected with flow cytometry. ROS scavenger (NAC) and p38-MAPK inhibitor (SB203580) were applied to resist the cytotoxic effect. Caspase 9 cleavage and p38 phosphorylation (P-p38) were quantified by western blot. Juglone as well as temozolomide (TMZ) were administrated in intracranial xenografts and MR scan was performed every week to evaluate the anti-tumor effect in vivo.

Results: Juglone could obviously inhibit the proliferation of TSCs in glioma by decreasing cell viability (P&thinsp;&lt;&thinsp;0.01) and inducing apoptosis (P&thinsp;&lt;&thinsp;0.01), which was accompanied by increased caspase 9 cleavage in a dose-dependent manner (P&thinsp;&lt;&thinsp;0.01). In the meantime, juglone could generate ROS significantly and increase p38 phosphorylation (P&thinsp;&lt;&thinsp;0.01). In addition, pretreatment with ROS scavenger or p38-MAPK inhibitor could reverse juglone-induced cytotoxicity (P&thinsp;&lt;&thinsp;0.01). More importantly, juglone could also suppress tumor growth in vivo and improve the survival of U87-bearing mice compared with control (P&thinsp;&lt;&thinsp;0.05), although TMZ seemed to have better effect.

Conclusions: Juglone could inhibit the growth of TSCs in gliomas through the activation of ROS-p38-MAPK pathway in vitro, and the anti-glioma effect was validated in vivo, which offers a potential therapeutic agent to gliomas.

No MeSH data available.