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Celastrol induces apoptosis of gastric cancer cells by miR-146a inhibition of NF-κB activity.

Sha M, Ye J, Zhang LX, Luan ZY, Chen YB - Cancer Cell Int. (2013)

Bottom Line: Finally, the effect of miR-146a on celastrol-induced anti-tumor activity was assessed using miR-146a inhibitor.Celastrol also reduced IκB phosphorylation, nuclear P65 protein levels and NF-κB activity.In this study, we demonstrated that the effect of celastrol on apoptosis is due to miR-146a inhibition of NF-κB activity.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Clinical medicine, Taizhou people's Hospital affiliated of Nantong University of medicine, 210 Yingchun, Taizhou, Jiangsu Province, 225300, China. tzrmyy5211@163.com.

ABSTRACT

Background: Celastrol, a plant triterpene, is known to play important role in inhibiting proliferation and inducing apoptosis of gastric cancer cells. In the present study, the mechanism of celastrol on gastric cancer cells apoptosis was examined.

Methods: We assessed effect of celastrol on NF-κB signaling pathway in gastric cancer cells using western blot and luciferase reporter assay. The real-time PCR was used to evaluate the effect of celastrol on miR-146a expression, and miR-146a mimic to evaluate whether over-expression of miR-146a can affect NF-κB activity. Finally, the effect of miR-146a on celastrol-induced anti-tumor activity was assessed using miR-146a inhibitor.

Results: Celastrol decreased gastric cancer cells viability in a dose-dependent. Celastrol also reduced IκB phosphorylation, nuclear P65 protein levels and NF-κB activity. Furthermore, Celastrol could increase miR-146a expression and up-regulation of miR-146a expression could suppress NF-κB activity. More important, down-regulation of miR-146a expression can reverse the effect of celastrol on NF-κB activity and apoptosis in gastric cancer cells.

Conclusions: In this study, we demonstrated that the effect of celastrol on apoptosis is due to miR-146a inhibition of NF-κB activity.

No MeSH data available.


Related in: MedlinePlus

Up-regulation of miR-146a expression can inhibit NF-κB activity in BGC-823, SGC-7901 and MGC-803 cells. The real-time PCR revealed that miR-146a mimic significantly increased the expression of miR-146a (A). Western blot showed that transfected miR-146a mimic decreased phosphorylation of IκB and nuclear p65 level (B). The luciferase reporter assay revealed that transfected miR-146a mimc significantly repressed NF-κB transcriptional activity (C). *P < 0.05, indicate significant differences from the respective control groups.
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Figure 3: Up-regulation of miR-146a expression can inhibit NF-κB activity in BGC-823, SGC-7901 and MGC-803 cells. The real-time PCR revealed that miR-146a mimic significantly increased the expression of miR-146a (A). Western blot showed that transfected miR-146a mimic decreased phosphorylation of IκB and nuclear p65 level (B). The luciferase reporter assay revealed that transfected miR-146a mimc significantly repressed NF-κB transcriptional activity (C). *P < 0.05, indicate significant differences from the respective control groups.

Mentions: To determine whether miR-146a can regulate signaling pathway in gastric cancer cells, we investigated the modulation of phosphorylation of IκB in cells transfected miR-146a mimic by western blot. The result of real-time PCR revealed that miR-146a mimic can significantly increase the expression of miR-146a in BGC-823, SGC-7901 and MGC-803 cells (P < 0.01) (Figure 3A), suggesting that miR-146a mimic is efficiently introduced into the cells and acts to up-regulate miR-146a expression. Furthermore, transfection of miR-146a mimic decreased phosphorylation of IκB and nuclear P65 as shown in Figure 3B.


Celastrol induces apoptosis of gastric cancer cells by miR-146a inhibition of NF-κB activity.

Sha M, Ye J, Zhang LX, Luan ZY, Chen YB - Cancer Cell Int. (2013)

Up-regulation of miR-146a expression can inhibit NF-κB activity in BGC-823, SGC-7901 and MGC-803 cells. The real-time PCR revealed that miR-146a mimic significantly increased the expression of miR-146a (A). Western blot showed that transfected miR-146a mimic decreased phosphorylation of IκB and nuclear p65 level (B). The luciferase reporter assay revealed that transfected miR-146a mimc significantly repressed NF-κB transcriptional activity (C). *P < 0.05, indicate significant differences from the respective control groups.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3672015&req=5

Figure 3: Up-regulation of miR-146a expression can inhibit NF-κB activity in BGC-823, SGC-7901 and MGC-803 cells. The real-time PCR revealed that miR-146a mimic significantly increased the expression of miR-146a (A). Western blot showed that transfected miR-146a mimic decreased phosphorylation of IκB and nuclear p65 level (B). The luciferase reporter assay revealed that transfected miR-146a mimc significantly repressed NF-κB transcriptional activity (C). *P < 0.05, indicate significant differences from the respective control groups.
Mentions: To determine whether miR-146a can regulate signaling pathway in gastric cancer cells, we investigated the modulation of phosphorylation of IκB in cells transfected miR-146a mimic by western blot. The result of real-time PCR revealed that miR-146a mimic can significantly increase the expression of miR-146a in BGC-823, SGC-7901 and MGC-803 cells (P < 0.01) (Figure 3A), suggesting that miR-146a mimic is efficiently introduced into the cells and acts to up-regulate miR-146a expression. Furthermore, transfection of miR-146a mimic decreased phosphorylation of IκB and nuclear P65 as shown in Figure 3B.

Bottom Line: Finally, the effect of miR-146a on celastrol-induced anti-tumor activity was assessed using miR-146a inhibitor.Celastrol also reduced IκB phosphorylation, nuclear P65 protein levels and NF-κB activity.In this study, we demonstrated that the effect of celastrol on apoptosis is due to miR-146a inhibition of NF-κB activity.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Clinical medicine, Taizhou people's Hospital affiliated of Nantong University of medicine, 210 Yingchun, Taizhou, Jiangsu Province, 225300, China. tzrmyy5211@163.com.

ABSTRACT

Background: Celastrol, a plant triterpene, is known to play important role in inhibiting proliferation and inducing apoptosis of gastric cancer cells. In the present study, the mechanism of celastrol on gastric cancer cells apoptosis was examined.

Methods: We assessed effect of celastrol on NF-κB signaling pathway in gastric cancer cells using western blot and luciferase reporter assay. The real-time PCR was used to evaluate the effect of celastrol on miR-146a expression, and miR-146a mimic to evaluate whether over-expression of miR-146a can affect NF-κB activity. Finally, the effect of miR-146a on celastrol-induced anti-tumor activity was assessed using miR-146a inhibitor.

Results: Celastrol decreased gastric cancer cells viability in a dose-dependent. Celastrol also reduced IκB phosphorylation, nuclear P65 protein levels and NF-κB activity. Furthermore, Celastrol could increase miR-146a expression and up-regulation of miR-146a expression could suppress NF-κB activity. More important, down-regulation of miR-146a expression can reverse the effect of celastrol on NF-κB activity and apoptosis in gastric cancer cells.

Conclusions: In this study, we demonstrated that the effect of celastrol on apoptosis is due to miR-146a inhibition of NF-κB activity.

No MeSH data available.


Related in: MedlinePlus