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A novel sLRP6E1E2 inhibits canonical Wnt signaling, epithelial-to-mesenchymal transition, and induces mitochondria-dependent apoptosis in lung cancer.

Lee JS, Hur MW, Lee SK, Choi WI, Kwon YG, Yun CO - PLoS ONE (2012)

Bottom Line: We designed a Wnt antagonist sLRP6E1E2, and generated a replication-incompetent adenovirus (Ad), dE1-k35/sLRP6E1E2, and a replication-competent oncolytic Ad, RdB-k35/sLRP6E1E2, both expressing sLRP6E1E2. sLRP6E1E2 prevented Wnt-mediated stabilization of cytoplasmic β-catenin, decreased Wnt/β-catenin signaling and cell proliferation via the mitogen-activated protein kinase, and phosphatidylinositol 3-kinase pathways. sLRP6E1E2 induced apoptosis, cytochrome c release, and increased cleavage of PARP and caspase-3. sLRP6E1E2 suppressed growth of the human lung tumor xenograft, and reduced motility and invasion of cancer cells.In addition, sLRP6E1E2 upregulated expression of epithelial marker genes, while sLRP6E1E2 downregulated mesenchymal marker genes.Taken together, sLRP6E1E2, by inhibiting interaction between Wnt and its receptor, suppressed Wnt-induced cell proliferation and epithelial-to-mesenchymal transition.

View Article: PubMed Central - PubMed

Affiliation: Brain Korea 21 Project for Medical Sciences, Yonsei Cancer Center, Yonsei University College of Medicine, Seoul, Korea.

ABSTRACT
Aberrant activation of the Wnt pathway contributes to human cancer progression. Antagonists that interfere with Wnt ligand/receptor interactions can be useful in cancer treatments. In this study, we evaluated the therapeutic potential of a soluble Wnt receptor decoy in cancer gene therapy. We designed a Wnt antagonist sLRP6E1E2, and generated a replication-incompetent adenovirus (Ad), dE1-k35/sLRP6E1E2, and a replication-competent oncolytic Ad, RdB-k35/sLRP6E1E2, both expressing sLRP6E1E2. sLRP6E1E2 prevented Wnt-mediated stabilization of cytoplasmic β-catenin, decreased Wnt/β-catenin signaling and cell proliferation via the mitogen-activated protein kinase, and phosphatidylinositol 3-kinase pathways. sLRP6E1E2 induced apoptosis, cytochrome c release, and increased cleavage of PARP and caspase-3. sLRP6E1E2 suppressed growth of the human lung tumor xenograft, and reduced motility and invasion of cancer cells. In addition, sLRP6E1E2 upregulated expression of epithelial marker genes, while sLRP6E1E2 downregulated mesenchymal marker genes. Taken together, sLRP6E1E2, by inhibiting interaction between Wnt and its receptor, suppressed Wnt-induced cell proliferation and epithelial-to-mesenchymal transition.

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Related in: MedlinePlus

Wnt3a treatment results in the disruption of cell-cell junctions and epithelial-to-mesenchymal transition in tumor cells.(a) H322 cells were treated with Wnt3a (100 ng/ml) for the indicated times, and morphology changes were observed by light microscopy. Original magnification, ×200. (b) E-cadherin, Vimentin, and β-catenin mRNA levels in H322 cells after Wnt3a treatment. (c) H322 cells were stained with DAPI (blue), TRITC-labeled actin (red), or anti E-cadherin (green) after 24 incubation with or without Wnt3a (100 ng/ml). Original magnification, ×630.
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pone-0036520-g006: Wnt3a treatment results in the disruption of cell-cell junctions and epithelial-to-mesenchymal transition in tumor cells.(a) H322 cells were treated with Wnt3a (100 ng/ml) for the indicated times, and morphology changes were observed by light microscopy. Original magnification, ×200. (b) E-cadherin, Vimentin, and β-catenin mRNA levels in H322 cells after Wnt3a treatment. (c) H322 cells were stained with DAPI (blue), TRITC-labeled actin (red), or anti E-cadherin (green) after 24 incubation with or without Wnt3a (100 ng/ml). Original magnification, ×630.

Mentions: EMT is an important process in tumor development, and the Wnt/β-catenin signal pathway may play an important role in this process. Therefore, we investigated whether Wnt3a could induce EMT in H322 cells. We found that cells became elongated and spindle-shaped 1 day after Wnt3a treatment, resembling the morphology of mesenchymal cells (Fig. 6A). We also observed increased expression of mesenchymal markers Vimentin and β-catenin with a concomitant decrease in epithelial marker E-cadherin (Fig. 6B). Immunofluorescence staining revealed that actin and E-cadherin levels were dramatically reduced in cell–cell contacts after Wnt3a treatment (Fig. 6C and Figure S5).


A novel sLRP6E1E2 inhibits canonical Wnt signaling, epithelial-to-mesenchymal transition, and induces mitochondria-dependent apoptosis in lung cancer.

Lee JS, Hur MW, Lee SK, Choi WI, Kwon YG, Yun CO - PLoS ONE (2012)

Wnt3a treatment results in the disruption of cell-cell junctions and epithelial-to-mesenchymal transition in tumor cells.(a) H322 cells were treated with Wnt3a (100 ng/ml) for the indicated times, and morphology changes were observed by light microscopy. Original magnification, ×200. (b) E-cadherin, Vimentin, and β-catenin mRNA levels in H322 cells after Wnt3a treatment. (c) H322 cells were stained with DAPI (blue), TRITC-labeled actin (red), or anti E-cadherin (green) after 24 incubation with or without Wnt3a (100 ng/ml). Original magnification, ×630.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3351461&req=5

pone-0036520-g006: Wnt3a treatment results in the disruption of cell-cell junctions and epithelial-to-mesenchymal transition in tumor cells.(a) H322 cells were treated with Wnt3a (100 ng/ml) for the indicated times, and morphology changes were observed by light microscopy. Original magnification, ×200. (b) E-cadherin, Vimentin, and β-catenin mRNA levels in H322 cells after Wnt3a treatment. (c) H322 cells were stained with DAPI (blue), TRITC-labeled actin (red), or anti E-cadherin (green) after 24 incubation with or without Wnt3a (100 ng/ml). Original magnification, ×630.
Mentions: EMT is an important process in tumor development, and the Wnt/β-catenin signal pathway may play an important role in this process. Therefore, we investigated whether Wnt3a could induce EMT in H322 cells. We found that cells became elongated and spindle-shaped 1 day after Wnt3a treatment, resembling the morphology of mesenchymal cells (Fig. 6A). We also observed increased expression of mesenchymal markers Vimentin and β-catenin with a concomitant decrease in epithelial marker E-cadherin (Fig. 6B). Immunofluorescence staining revealed that actin and E-cadherin levels were dramatically reduced in cell–cell contacts after Wnt3a treatment (Fig. 6C and Figure S5).

Bottom Line: We designed a Wnt antagonist sLRP6E1E2, and generated a replication-incompetent adenovirus (Ad), dE1-k35/sLRP6E1E2, and a replication-competent oncolytic Ad, RdB-k35/sLRP6E1E2, both expressing sLRP6E1E2. sLRP6E1E2 prevented Wnt-mediated stabilization of cytoplasmic β-catenin, decreased Wnt/β-catenin signaling and cell proliferation via the mitogen-activated protein kinase, and phosphatidylinositol 3-kinase pathways. sLRP6E1E2 induced apoptosis, cytochrome c release, and increased cleavage of PARP and caspase-3. sLRP6E1E2 suppressed growth of the human lung tumor xenograft, and reduced motility and invasion of cancer cells.In addition, sLRP6E1E2 upregulated expression of epithelial marker genes, while sLRP6E1E2 downregulated mesenchymal marker genes.Taken together, sLRP6E1E2, by inhibiting interaction between Wnt and its receptor, suppressed Wnt-induced cell proliferation and epithelial-to-mesenchymal transition.

View Article: PubMed Central - PubMed

Affiliation: Brain Korea 21 Project for Medical Sciences, Yonsei Cancer Center, Yonsei University College of Medicine, Seoul, Korea.

ABSTRACT
Aberrant activation of the Wnt pathway contributes to human cancer progression. Antagonists that interfere with Wnt ligand/receptor interactions can be useful in cancer treatments. In this study, we evaluated the therapeutic potential of a soluble Wnt receptor decoy in cancer gene therapy. We designed a Wnt antagonist sLRP6E1E2, and generated a replication-incompetent adenovirus (Ad), dE1-k35/sLRP6E1E2, and a replication-competent oncolytic Ad, RdB-k35/sLRP6E1E2, both expressing sLRP6E1E2. sLRP6E1E2 prevented Wnt-mediated stabilization of cytoplasmic β-catenin, decreased Wnt/β-catenin signaling and cell proliferation via the mitogen-activated protein kinase, and phosphatidylinositol 3-kinase pathways. sLRP6E1E2 induced apoptosis, cytochrome c release, and increased cleavage of PARP and caspase-3. sLRP6E1E2 suppressed growth of the human lung tumor xenograft, and reduced motility and invasion of cancer cells. In addition, sLRP6E1E2 upregulated expression of epithelial marker genes, while sLRP6E1E2 downregulated mesenchymal marker genes. Taken together, sLRP6E1E2, by inhibiting interaction between Wnt and its receptor, suppressed Wnt-induced cell proliferation and epithelial-to-mesenchymal transition.

Show MeSH
Related in: MedlinePlus