Limits...
WNT5A promotes stemness characteristics in nasopharyngeal carcinoma cells leading to metastasis and tumorigenesis.

Qin L, Yin YT, Zheng FJ, Peng LX, Yang CF, Bao YN, Liang YY, Li XJ, Xiang YQ, Sun R, Li AH, Zou RH, Pei XQ, Huang BJ, Kang TB, Liao DF, Zeng YX, Williams BO, Qian CN - Oncotarget (2015)

Bottom Line: When elevated expression of WNT5A coincided with the elevated expression of vimentin in the primary NPC, the patients had a poorer prognosis.Among major signaling pathways, protein kinase C (PKC) signaling was activated by WNT5A in NPC cells.A positive feedback loop between WNT5A and phospho-PKC to promote EMT was also revealed.

View Article: PubMed Central - PubMed

Affiliation: Sun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou, China.

ABSTRACT
Nasopharyngeal carcinoma (NPC) has the highest metastasis rate among head and neck cancers with unclear mechanism. WNT5A belongs to the WNT family of cysteine-rich secreted glycoproteins. Our previous high-throughput gene expression profiling revealed that WNT5A was up-regulated in highly metastatic cells. In the present study, we first confirmed the elevated expression of WNT5A in metastatic NPC tissues at both the mRNA and protein levels. We then found that WNT5A promoted epithelial-mesenchymal transition (EMT) in NPC cells, induced the accumulation of CD24-CD44+ cells and side population, which are believed to be cancer stem cell characteristics. Moreover, WNT5A promoted the migration and invasion of NPC cells in vitro, while in vivo treatment with recombinant WNT5A promoted lung metastasis. Knocking down WNT5A diminished NPC tumorigenesis in vivo. When elevated expression of WNT5A coincided with the elevated expression of vimentin in the primary NPC, the patients had a poorer prognosis. Among major signaling pathways, protein kinase C (PKC) signaling was activated by WNT5A in NPC cells. A positive feedback loop between WNT5A and phospho-PKC to promote EMT was also revealed. Taken together, these data suggest that WNT5A is an important molecule in promoting stem cell characteristics in NPC, leading to tumorigenesis and metastasis.

No MeSH data available.


Related in: MedlinePlus

WNT5A activates PKC signalingA positive loop between WNT5A and phospho-PKC promotes EMT in NPC cells. A, By using SUNE-1 cells with WNT5A over-expression as well as 5-8F cells with WNT5A knocked-down, several major signaling pathways were examined, including PKC, ERK, AKT, and JNK pathways. Only phosphorylation of PKC was altered by WNT5A expression in the cells. B, Over-expression or knocking down of WNT5A did not alter the protein level of β-catenin in the nucleus, indicating that the classic β-catenin signaling pathway could not be activated by WNT5A in NPC cells. C, Over-expression of WNT5A increased phospho-PKC level, while knocking down WNT5A resulted in reduction of phosphorylated PKC. D, PKC activator PMA could successfully induce phosphorylation of PKC in S26 cells. E, Activation of PKC by PMA in S26 cells could further up-regulate WNT5A level and subsequently increase mesenchymal marker Snail and decrease epithelial marker E-cadherin. F, PKC inhibitor GF10923X could decrease WNT5A level in S18 cells and subsequently reduce Snail and gain E-cadherin.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4496352&req=5

Figure 5: WNT5A activates PKC signalingA positive loop between WNT5A and phospho-PKC promotes EMT in NPC cells. A, By using SUNE-1 cells with WNT5A over-expression as well as 5-8F cells with WNT5A knocked-down, several major signaling pathways were examined, including PKC, ERK, AKT, and JNK pathways. Only phosphorylation of PKC was altered by WNT5A expression in the cells. B, Over-expression or knocking down of WNT5A did not alter the protein level of β-catenin in the nucleus, indicating that the classic β-catenin signaling pathway could not be activated by WNT5A in NPC cells. C, Over-expression of WNT5A increased phospho-PKC level, while knocking down WNT5A resulted in reduction of phosphorylated PKC. D, PKC activator PMA could successfully induce phosphorylation of PKC in S26 cells. E, Activation of PKC by PMA in S26 cells could further up-regulate WNT5A level and subsequently increase mesenchymal marker Snail and decrease epithelial marker E-cadherin. F, PKC inhibitor GF10923X could decrease WNT5A level in S18 cells and subsequently reduce Snail and gain E-cadherin.

Mentions: We further examined the roles of WNT5A in several major signaling pathways underlying tumor progression, including PKC, ERK, AKT, and JNK pathways. PKC signaling was the one activated by WNT5A in NPC cells (Figure 5A). However, the classical β-catenin signaling was not activated by WNT5A (Figure 5B). We further tested whether the positive feedback loop between WNT5A and phospho-PKC could regulate EMT in NPC cells. First, we found that stably knocking down WNT5A mRNA using shRNAs in high-metastasis S18 cells significantly diminished PKC phosphorylation, and overexpression of WNT5A up-regulated phospho-PKC in low-metastasis S26 cells (Figure 5C). Activating PKC using the PKC activator PMA in S26 cells (Figure 5D) induced the accumulation of WNT5A and subsequently induced the up-regulation of the mesenchymal marker Snail and a reduction in the epithelial marker E-cadherin (Figure 5E). Inhibiting PKC phosphorylation in S18 cells using GF10923X reduced the phospho-PKC level and down-regulated the WNT5A level with a consequent reduction in Snail and an increase in E-cadherin (Figure 5F). Taken together, the positive feedback loop between phospho-PKC and WNT5A triggered EMT in NPC cells (Figure 6).


WNT5A promotes stemness characteristics in nasopharyngeal carcinoma cells leading to metastasis and tumorigenesis.

Qin L, Yin YT, Zheng FJ, Peng LX, Yang CF, Bao YN, Liang YY, Li XJ, Xiang YQ, Sun R, Li AH, Zou RH, Pei XQ, Huang BJ, Kang TB, Liao DF, Zeng YX, Williams BO, Qian CN - Oncotarget (2015)

WNT5A activates PKC signalingA positive loop between WNT5A and phospho-PKC promotes EMT in NPC cells. A, By using SUNE-1 cells with WNT5A over-expression as well as 5-8F cells with WNT5A knocked-down, several major signaling pathways were examined, including PKC, ERK, AKT, and JNK pathways. Only phosphorylation of PKC was altered by WNT5A expression in the cells. B, Over-expression or knocking down of WNT5A did not alter the protein level of β-catenin in the nucleus, indicating that the classic β-catenin signaling pathway could not be activated by WNT5A in NPC cells. C, Over-expression of WNT5A increased phospho-PKC level, while knocking down WNT5A resulted in reduction of phosphorylated PKC. D, PKC activator PMA could successfully induce phosphorylation of PKC in S26 cells. E, Activation of PKC by PMA in S26 cells could further up-regulate WNT5A level and subsequently increase mesenchymal marker Snail and decrease epithelial marker E-cadherin. F, PKC inhibitor GF10923X could decrease WNT5A level in S18 cells and subsequently reduce Snail and gain E-cadherin.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4496352&req=5

Figure 5: WNT5A activates PKC signalingA positive loop between WNT5A and phospho-PKC promotes EMT in NPC cells. A, By using SUNE-1 cells with WNT5A over-expression as well as 5-8F cells with WNT5A knocked-down, several major signaling pathways were examined, including PKC, ERK, AKT, and JNK pathways. Only phosphorylation of PKC was altered by WNT5A expression in the cells. B, Over-expression or knocking down of WNT5A did not alter the protein level of β-catenin in the nucleus, indicating that the classic β-catenin signaling pathway could not be activated by WNT5A in NPC cells. C, Over-expression of WNT5A increased phospho-PKC level, while knocking down WNT5A resulted in reduction of phosphorylated PKC. D, PKC activator PMA could successfully induce phosphorylation of PKC in S26 cells. E, Activation of PKC by PMA in S26 cells could further up-regulate WNT5A level and subsequently increase mesenchymal marker Snail and decrease epithelial marker E-cadherin. F, PKC inhibitor GF10923X could decrease WNT5A level in S18 cells and subsequently reduce Snail and gain E-cadherin.
Mentions: We further examined the roles of WNT5A in several major signaling pathways underlying tumor progression, including PKC, ERK, AKT, and JNK pathways. PKC signaling was the one activated by WNT5A in NPC cells (Figure 5A). However, the classical β-catenin signaling was not activated by WNT5A (Figure 5B). We further tested whether the positive feedback loop between WNT5A and phospho-PKC could regulate EMT in NPC cells. First, we found that stably knocking down WNT5A mRNA using shRNAs in high-metastasis S18 cells significantly diminished PKC phosphorylation, and overexpression of WNT5A up-regulated phospho-PKC in low-metastasis S26 cells (Figure 5C). Activating PKC using the PKC activator PMA in S26 cells (Figure 5D) induced the accumulation of WNT5A and subsequently induced the up-regulation of the mesenchymal marker Snail and a reduction in the epithelial marker E-cadherin (Figure 5E). Inhibiting PKC phosphorylation in S18 cells using GF10923X reduced the phospho-PKC level and down-regulated the WNT5A level with a consequent reduction in Snail and an increase in E-cadherin (Figure 5F). Taken together, the positive feedback loop between phospho-PKC and WNT5A triggered EMT in NPC cells (Figure 6).

Bottom Line: When elevated expression of WNT5A coincided with the elevated expression of vimentin in the primary NPC, the patients had a poorer prognosis.Among major signaling pathways, protein kinase C (PKC) signaling was activated by WNT5A in NPC cells.A positive feedback loop between WNT5A and phospho-PKC to promote EMT was also revealed.

View Article: PubMed Central - PubMed

Affiliation: Sun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou, China.

ABSTRACT
Nasopharyngeal carcinoma (NPC) has the highest metastasis rate among head and neck cancers with unclear mechanism. WNT5A belongs to the WNT family of cysteine-rich secreted glycoproteins. Our previous high-throughput gene expression profiling revealed that WNT5A was up-regulated in highly metastatic cells. In the present study, we first confirmed the elevated expression of WNT5A in metastatic NPC tissues at both the mRNA and protein levels. We then found that WNT5A promoted epithelial-mesenchymal transition (EMT) in NPC cells, induced the accumulation of CD24-CD44+ cells and side population, which are believed to be cancer stem cell characteristics. Moreover, WNT5A promoted the migration and invasion of NPC cells in vitro, while in vivo treatment with recombinant WNT5A promoted lung metastasis. Knocking down WNT5A diminished NPC tumorigenesis in vivo. When elevated expression of WNT5A coincided with the elevated expression of vimentin in the primary NPC, the patients had a poorer prognosis. Among major signaling pathways, protein kinase C (PKC) signaling was activated by WNT5A in NPC cells. A positive feedback loop between WNT5A and phospho-PKC to promote EMT was also revealed. Taken together, these data suggest that WNT5A is an important molecule in promoting stem cell characteristics in NPC, leading to tumorigenesis and metastasis.

No MeSH data available.


Related in: MedlinePlus