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Inhibition of NF-kappa B pathway leads to deregulation of epithelial-mesenchymal transition and neural invasion in pancreatic cancer

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ABSTRACT

NF-κB plays an essential role in the initiation and progression of pancreatic cancer and specifically mediates the induction of epithelial-mesenchymal transition and invasiveness. In this study we demonstrate the importance of activated NF-κB signaling in EMT induction, lymphovascular metastasis, and neural invasion. Modulation of NF-κB activity was accomplished through the specific NF-κB inhibitor (BAY 11-7085), triptolide, and Minnelide treatment, as well as overexpression of IKBα repressor and IKK activator plasmids. In the classical lymphovascular metastatic cascade, inhibition of NF-κB decreased the expression of several EMT transcription factors (SNAI1, SNAI2, ZEB1) and mesenchymal markers (VIM and CDH2) and decreased in vitro invasion, which was rescued by IKK activation. This was further demonstrated in vivo via BAY 11-7085 treatment in a orthotopic model of pancreatic cancer. In vivo NF-κB inhibition decreased tumor volume; decreased tumor EMT gene expression, while restoring cell-cell junctions; and decreased overall metastasis. Furthermore, we demonstrate the importance of active NF-κB signaling in neural invasion. Triptolide treatment inhibits NGF mediated and neural-tumor co-culture in vitro invasion and dorsal root ganglia (DRG) neural outgrowth through a disruption in tumor-neural cross talk. In vivo, Minnelide treatment decreased neurotrophin expression, nerve density, and sciatic nerve invasion. Taken together, this study demonstrates the importance of NF-κB signaling in the progression of pancreatic cancer through the modulation of EMT induction, lymphovascular invasion, and neural invasion.

No MeSH data available.


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Inhibition of NF-κB inhibits EMT and metastasis in vivo: BAY 11-7085 treatment in a orthotopic MIA PaCa-2 model decreases (a) tumor weight; (b) tumor NF-κB activity; (c) tumor EMT gene expression; and (d) (e) metastasis to distant sites. Each bar is representative of three or more independent experiments; error bars are represented in SEM; and the asterisk (*) indicates a p value < 0.05.
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Figure 4: Inhibition of NF-κB inhibits EMT and metastasis in vivo: BAY 11-7085 treatment in a orthotopic MIA PaCa-2 model decreases (a) tumor weight; (b) tumor NF-κB activity; (c) tumor EMT gene expression; and (d) (e) metastasis to distant sites. Each bar is representative of three or more independent experiments; error bars are represented in SEM; and the asterisk (*) indicates a p value < 0.05.

Mentions: To determine whether the EMT inhibition and invasion which was seen in vitro translates in vivo, we used BAY 11-7085 treatment via intraperitoneal injection to inhibit NF-κB signaling in an orthotopic mouse model of pancreatic cancer. Treatment for 3 weeks resulted in a decrease in primary tumor weight (0.561g ± 0.078 in the control group versus 0.250g ± 0.052 in the BAY 11-7085 treatment group) (Figure 4a). Tumor NF-κB activity was determined by p50 binding ELISA, in which BAY 11-7085 treated tumors displayed decreased p50 binding (9657 RLU/μg protein ± 586.6) as compared to DMSO control (7248 RLU/μg protein ± 538.3) (Figure 4b). BAY 11-7085 treated tumors also exhibited decreased gene expression of EMT transcription factors (SNAI2 0.745 fold ± 0.070 and TWIST1 0.737 fold ± 0.164) and mesenchymal marker (N-Cadherin 0.377 fold ± 0.196); increased E-cadherin expression (2.392 fold ± 0.265); and decreased MMP7 gene expression (0.002 fold ± 0.001) (Figure 4c).


Inhibition of NF-kappa B pathway leads to deregulation of epithelial-mesenchymal transition and neural invasion in pancreatic cancer
Inhibition of NF-κB inhibits EMT and metastasis in vivo: BAY 11-7085 treatment in a orthotopic MIA PaCa-2 model decreases (a) tumor weight; (b) tumor NF-κB activity; (c) tumor EMT gene expression; and (d) (e) metastasis to distant sites. Each bar is representative of three or more independent experiments; error bars are represented in SEM; and the asterisk (*) indicates a p value < 0.05.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC5121017&req=5

Figure 4: Inhibition of NF-κB inhibits EMT and metastasis in vivo: BAY 11-7085 treatment in a orthotopic MIA PaCa-2 model decreases (a) tumor weight; (b) tumor NF-κB activity; (c) tumor EMT gene expression; and (d) (e) metastasis to distant sites. Each bar is representative of three or more independent experiments; error bars are represented in SEM; and the asterisk (*) indicates a p value < 0.05.
Mentions: To determine whether the EMT inhibition and invasion which was seen in vitro translates in vivo, we used BAY 11-7085 treatment via intraperitoneal injection to inhibit NF-κB signaling in an orthotopic mouse model of pancreatic cancer. Treatment for 3 weeks resulted in a decrease in primary tumor weight (0.561g ± 0.078 in the control group versus 0.250g ± 0.052 in the BAY 11-7085 treatment group) (Figure 4a). Tumor NF-κB activity was determined by p50 binding ELISA, in which BAY 11-7085 treated tumors displayed decreased p50 binding (9657 RLU/μg protein ± 586.6) as compared to DMSO control (7248 RLU/μg protein ± 538.3) (Figure 4b). BAY 11-7085 treated tumors also exhibited decreased gene expression of EMT transcription factors (SNAI2 0.745 fold ± 0.070 and TWIST1 0.737 fold ± 0.164) and mesenchymal marker (N-Cadherin 0.377 fold ± 0.196); increased E-cadherin expression (2.392 fold ± 0.265); and decreased MMP7 gene expression (0.002 fold ± 0.001) (Figure 4c).

View Article: PubMed Central - PubMed

ABSTRACT

NF-&kappa;B plays an essential role in the initiation and progression of pancreatic cancer and specifically mediates the induction of epithelial-mesenchymal transition and invasiveness. In this study we demonstrate the importance of activated NF-&kappa;B signaling in EMT induction, lymphovascular metastasis, and neural invasion. Modulation of NF-&kappa;B activity was accomplished through the specific NF-&kappa;B inhibitor (BAY 11-7085), triptolide, and Minnelide treatment, as well as overexpression of IKB&alpha; repressor and IKK activator plasmids. In the classical lymphovascular metastatic cascade, inhibition of NF-&kappa;B decreased the expression of several EMT transcription factors (SNAI1, SNAI2, ZEB1) and mesenchymal markers (VIM and CDH2) and decreased in vitro invasion, which was rescued by IKK activation. This was further demonstrated in vivo via BAY 11-7085 treatment in a orthotopic model of pancreatic cancer. In vivo NF-&kappa;B inhibition decreased tumor volume; decreased tumor EMT gene expression, while restoring cell-cell junctions; and decreased overall metastasis. Furthermore, we demonstrate the importance of active NF-&kappa;B signaling in neural invasion. Triptolide treatment inhibits NGF mediated and neural-tumor co-culture in vitro invasion and dorsal root ganglia (DRG) neural outgrowth through a disruption in tumor-neural cross talk. In vivo, Minnelide treatment decreased neurotrophin expression, nerve density, and sciatic nerve invasion. Taken together, this study demonstrates the importance of NF-&kappa;B signaling in the progression of pancreatic cancer through the modulation of EMT induction, lymphovascular invasion, and neural invasion.

No MeSH data available.


Related in: MedlinePlus