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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

Down-expression of miR-146a can reverse the effect of celastrol on NF-κB activity and apoptosis. The real-time PCR revealed that miR-146a inhibitor can significantly decrease the expression of miR-146a in BGC-823, SGC-7901 and MGC-803 cells (A). After transfected with 100 nmol/L of miR-146a inhibitor, the cells were treated with 2 μM celastrol for 6 h. miR-146a inhibitor significantly increased NF-κB transcriptional activity in BGC-823, SGC-7901 and MGC-803 cells after celastrol treatment (B). Flow-cytometric analysis showed that miR-146a inhibitor significantly decreased the apoptosis of BGC-823, SGC-7901 and MGC-803 cells induced with celastrol (C). After transfected with 100 nmol/L of miR-146a inhibitor, the cells were treated with 2 μM celastrol for 72 h. MTT revealed that miR-146a inhibitor significantly decreased the effect of celastrol on inhibition of BGC-823, SGC-7901 and MGC-803 cells growth (D). *P < 0.05, indicate significant differences from the respective control groups.
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Figure 4: Down-expression of miR-146a can reverse the effect of celastrol on NF-κB activity and apoptosis. The real-time PCR revealed that miR-146a inhibitor can significantly decrease the expression of miR-146a in BGC-823, SGC-7901 and MGC-803 cells (A). After transfected with 100 nmol/L of miR-146a inhibitor, the cells were treated with 2 μM celastrol for 6 h. miR-146a inhibitor significantly increased NF-κB transcriptional activity in BGC-823, SGC-7901 and MGC-803 cells after celastrol treatment (B). Flow-cytometric analysis showed that miR-146a inhibitor significantly decreased the apoptosis of BGC-823, SGC-7901 and MGC-803 cells induced with celastrol (C). After transfected with 100 nmol/L of miR-146a inhibitor, the cells were treated with 2 μM celastrol for 72 h. MTT revealed that miR-146a inhibitor significantly decreased the effect of celastrol on inhibition of BGC-823, SGC-7901 and MGC-803 cells growth (D). *P < 0.05, indicate significant differences from the respective control groups.

Mentions: In order to evaluate the role of miR-146a in the effect of celastrol on inhibition of BGC-823, SGC-7901 and MGC-803 cells apoptosis, we treated cells with celastrol after transfected with miR-146a inhibitor. As shown in Figure 4A, miR-146a inhibitor can significantly decrease the expression of miR-146a in BGC-823, SGC-7901 and MGC-803 cells (P < 0.01). Interestingly, we found that celastrol treatment inhibited transcriptional activity of NF-κB, which can be restored by inhibition of miR-146a expression (Figure 4B). In addition, the number of cells that underwent apoptosis significantly increased after treated with 2 μM celastrol. Moreover, the celastrol induced apoptosis was attenuated by miR-146a inhibitor in BGC-823, SGC-7901 and MGC-803 cells (Figure 4C). Furthermore, BGC-823, SGC-7901 and MGC-803 cells growth was increased in celastrol-treated cells after transfected miR-146a inhibitor (Figure 4D).


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)

Down-expression of miR-146a can reverse the effect of celastrol on NF-κB activity and apoptosis. The real-time PCR revealed that miR-146a inhibitor can significantly decrease the expression of miR-146a in BGC-823, SGC-7901 and MGC-803 cells (A). After transfected with 100 nmol/L of miR-146a inhibitor, the cells were treated with 2 μM celastrol for 6 h. miR-146a inhibitor significantly increased NF-κB transcriptional activity in BGC-823, SGC-7901 and MGC-803 cells after celastrol treatment (B). Flow-cytometric analysis showed that miR-146a inhibitor significantly decreased the apoptosis of BGC-823, SGC-7901 and MGC-803 cells induced with celastrol (C). After transfected with 100 nmol/L of miR-146a inhibitor, the cells were treated with 2 μM celastrol for 72 h. MTT revealed that miR-146a inhibitor significantly decreased the effect of celastrol on inhibition of BGC-823, SGC-7901 and MGC-803 cells growth (D). *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 4: Down-expression of miR-146a can reverse the effect of celastrol on NF-κB activity and apoptosis. The real-time PCR revealed that miR-146a inhibitor can significantly decrease the expression of miR-146a in BGC-823, SGC-7901 and MGC-803 cells (A). After transfected with 100 nmol/L of miR-146a inhibitor, the cells were treated with 2 μM celastrol for 6 h. miR-146a inhibitor significantly increased NF-κB transcriptional activity in BGC-823, SGC-7901 and MGC-803 cells after celastrol treatment (B). Flow-cytometric analysis showed that miR-146a inhibitor significantly decreased the apoptosis of BGC-823, SGC-7901 and MGC-803 cells induced with celastrol (C). After transfected with 100 nmol/L of miR-146a inhibitor, the cells were treated with 2 μM celastrol for 72 h. MTT revealed that miR-146a inhibitor significantly decreased the effect of celastrol on inhibition of BGC-823, SGC-7901 and MGC-803 cells growth (D). *P < 0.05, indicate significant differences from the respective control groups.
Mentions: In order to evaluate the role of miR-146a in the effect of celastrol on inhibition of BGC-823, SGC-7901 and MGC-803 cells apoptosis, we treated cells with celastrol after transfected with miR-146a inhibitor. As shown in Figure 4A, miR-146a inhibitor can significantly decrease the expression of miR-146a in BGC-823, SGC-7901 and MGC-803 cells (P < 0.01). Interestingly, we found that celastrol treatment inhibited transcriptional activity of NF-κB, which can be restored by inhibition of miR-146a expression (Figure 4B). In addition, the number of cells that underwent apoptosis significantly increased after treated with 2 μM celastrol. Moreover, the celastrol induced apoptosis was attenuated by miR-146a inhibitor in BGC-823, SGC-7901 and MGC-803 cells (Figure 4C). Furthermore, BGC-823, SGC-7901 and MGC-803 cells growth was increased in celastrol-treated cells after transfected miR-146a inhibitor (Figure 4D).

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