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A novel role for Wnt/Ca2+ signaling in actin cytoskeleton remodeling and cell motility in prostate cancer.

Wang Q, Symes AJ, Kane CA, Freeman A, Nariculam J, Munson P, Thrasivoulou C, Masters JR, Ahmed A - PLoS ONE (2010)

Bottom Line: Wnt signaling is a critical regulatory pathway in development and disease.Live cell imaging showed that a functional consequence of CaMKII inhibition was 80% decrease in wound healing capacity and reduced cell motility in cancer cells.We propose that non-canonical Wnt/Ca(2+) signaling via CaMKII acts as a novel regulator of structural plasticity and cell motility in prostate cancer.

View Article: PubMed Central - PubMed

Affiliation: Prostate Cancer Research Centre and Division of Surgery, University College London, London, United Kingdom.

ABSTRACT
Wnt signaling is a critical regulatory pathway in development and disease. Very little is known about the mechanisms of Wnt signaling in prostate cancer, a leading cause of death in men. A quantitative analysis of the expression of Wnt5A protein in human tissue arrays, containing 600 prostate tissue cores, showed >50% increase in malignant compared to benign cores (p<0.0001). In a matched pair of prostate cancer and normal cell line, expression of Wnt5A protein was also increased. Calcium waves were induced in prostate cells in response to Wnt5A with a 3 fold increase in Flou-4 intensity. The activity of Ca(2+)/calmodulin dependent protein kinase (CaMKII), a transducer of the non-canonical Wnt/Ca(2+) signaling, increased by 8 fold in cancer cells; no change was observed in beta-catenin expression, known to activate the canonical Wnt/beta-catenin pathway. Mining of publicly available human prostate cancer oligoarray datasets revealed that the expression of numerous genes (e.g., CCND1, CD44) under the control of beta-catenin transcription is down-regulated. Confocal and quantitative electron microscopy showed that specific inhibition of CaMKII in cancer cells causes remodeling of the actin cytoskeleton, irregular wound edges and loose intercellular architecture and a 6 and 8 fold increase in the frequency and length of filopodia, respectively. Conversely, untreated normal prostate cells showed an irregular wound edge and loose intercellular architecture; incubation of normal prostate cells with recombinant Wnt5A protein induced actin remodeling with a regular wound edge and increased wound healing capacity. Live cell imaging showed that a functional consequence of CaMKII inhibition was 80% decrease in wound healing capacity and reduced cell motility in cancer cells. We propose that non-canonical Wnt/Ca(2+) signaling via CaMKII acts as a novel regulator of structural plasticity and cell motility in prostate cancer.

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Related in: MedlinePlus

CaMKII activity in prostate cells.The phosphotransferase activity of CaMKII in cell lysates was measured by using a [γ-32P] ATP based CaMKII assay (Upstate, UK) with Ca2+ (hatched bars) or without Ca2+ (hollow bars). Inset: Ca2+-dependent CaMKII activity was calculated as described in Materials and Methods. Values are mean ± SE from 3 experiments. NP is1542-NPTX, CP is1542-CP3TX, DU is DU145 and PC is PC3 prostate cell line.
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pone-0010456-g004: CaMKII activity in prostate cells.The phosphotransferase activity of CaMKII in cell lysates was measured by using a [γ-32P] ATP based CaMKII assay (Upstate, UK) with Ca2+ (hatched bars) or without Ca2+ (hollow bars). Inset: Ca2+-dependent CaMKII activity was calculated as described in Materials and Methods. Values are mean ± SE from 3 experiments. NP is1542-NPTX, CP is1542-CP3TX, DU is DU145 and PC is PC3 prostate cell line.

Mentions: CamKII is a major transducer of Wnt/Ca2+ signaling. In all prostate cell lines CaMKII enzyme activity was Ca2+ dependent, least in 1542-NPTX, greater in 1542-CP3TX and DU145 and pronounced in PC3 cell line (Fig. 4). There was a 4 and 8 fold increase in the Ca2+-dependent CaMKII activity in1542-NPTX and 1542-CP3TX cells (Fig. 4), respectively. More importantly, the Ca2+ dependent activity of CaMKII was increased by ∼4 fold in 1542-CP3TX compared to 1542-NPTX (inset Fig. 4). These results indicate an increase in the activity of CaMKII in cancer cells compared to normal cells.


A novel role for Wnt/Ca2+ signaling in actin cytoskeleton remodeling and cell motility in prostate cancer.

Wang Q, Symes AJ, Kane CA, Freeman A, Nariculam J, Munson P, Thrasivoulou C, Masters JR, Ahmed A - PLoS ONE (2010)

CaMKII activity in prostate cells.The phosphotransferase activity of CaMKII in cell lysates was measured by using a [γ-32P] ATP based CaMKII assay (Upstate, UK) with Ca2+ (hatched bars) or without Ca2+ (hollow bars). Inset: Ca2+-dependent CaMKII activity was calculated as described in Materials and Methods. Values are mean ± SE from 3 experiments. NP is1542-NPTX, CP is1542-CP3TX, DU is DU145 and PC is PC3 prostate cell line.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2864254&req=5

pone-0010456-g004: CaMKII activity in prostate cells.The phosphotransferase activity of CaMKII in cell lysates was measured by using a [γ-32P] ATP based CaMKII assay (Upstate, UK) with Ca2+ (hatched bars) or without Ca2+ (hollow bars). Inset: Ca2+-dependent CaMKII activity was calculated as described in Materials and Methods. Values are mean ± SE from 3 experiments. NP is1542-NPTX, CP is1542-CP3TX, DU is DU145 and PC is PC3 prostate cell line.
Mentions: CamKII is a major transducer of Wnt/Ca2+ signaling. In all prostate cell lines CaMKII enzyme activity was Ca2+ dependent, least in 1542-NPTX, greater in 1542-CP3TX and DU145 and pronounced in PC3 cell line (Fig. 4). There was a 4 and 8 fold increase in the Ca2+-dependent CaMKII activity in1542-NPTX and 1542-CP3TX cells (Fig. 4), respectively. More importantly, the Ca2+ dependent activity of CaMKII was increased by ∼4 fold in 1542-CP3TX compared to 1542-NPTX (inset Fig. 4). These results indicate an increase in the activity of CaMKII in cancer cells compared to normal cells.

Bottom Line: Wnt signaling is a critical regulatory pathway in development and disease.Live cell imaging showed that a functional consequence of CaMKII inhibition was 80% decrease in wound healing capacity and reduced cell motility in cancer cells.We propose that non-canonical Wnt/Ca(2+) signaling via CaMKII acts as a novel regulator of structural plasticity and cell motility in prostate cancer.

View Article: PubMed Central - PubMed

Affiliation: Prostate Cancer Research Centre and Division of Surgery, University College London, London, United Kingdom.

ABSTRACT
Wnt signaling is a critical regulatory pathway in development and disease. Very little is known about the mechanisms of Wnt signaling in prostate cancer, a leading cause of death in men. A quantitative analysis of the expression of Wnt5A protein in human tissue arrays, containing 600 prostate tissue cores, showed >50% increase in malignant compared to benign cores (p<0.0001). In a matched pair of prostate cancer and normal cell line, expression of Wnt5A protein was also increased. Calcium waves were induced in prostate cells in response to Wnt5A with a 3 fold increase in Flou-4 intensity. The activity of Ca(2+)/calmodulin dependent protein kinase (CaMKII), a transducer of the non-canonical Wnt/Ca(2+) signaling, increased by 8 fold in cancer cells; no change was observed in beta-catenin expression, known to activate the canonical Wnt/beta-catenin pathway. Mining of publicly available human prostate cancer oligoarray datasets revealed that the expression of numerous genes (e.g., CCND1, CD44) under the control of beta-catenin transcription is down-regulated. Confocal and quantitative electron microscopy showed that specific inhibition of CaMKII in cancer cells causes remodeling of the actin cytoskeleton, irregular wound edges and loose intercellular architecture and a 6 and 8 fold increase in the frequency and length of filopodia, respectively. Conversely, untreated normal prostate cells showed an irregular wound edge and loose intercellular architecture; incubation of normal prostate cells with recombinant Wnt5A protein induced actin remodeling with a regular wound edge and increased wound healing capacity. Live cell imaging showed that a functional consequence of CaMKII inhibition was 80% decrease in wound healing capacity and reduced cell motility in cancer cells. We propose that non-canonical Wnt/Ca(2+) signaling via CaMKII acts as a novel regulator of structural plasticity and cell motility in prostate cancer.

Show MeSH
Related in: MedlinePlus