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Phosphorylation of calcium/calmodulin-stimulated protein kinase II at T286 enhances invasion and migration of human breast cancer cells

View Article: PubMed Central - PubMed

ABSTRACT

Calcium/calmodulin-stimulated protein kinase II (CaMKII) is a multi-functional kinase that controls a range of cellular functions, including proliferation, differentiation and apoptosis. The biological properties of CaMKII are regulated by multi-site phosphorylation. However, the role that CaMKII phosphorylation plays in cancer cell metastasis has not been examined. We demonstrate herein that CaMKII expression and phosphorylation at T286 is increased in breast cancer when compared to normal breast tissue, and that increased CAMK2 mRNA is associated with poor breast cancer patient prognosis (worse overall and distant metastasis free survival). Additionally, we show that overexpression of WT, T286D and T286V forms of CaMKII in MDA-MB-231 and MCF-7 breast cancer cells increases invasion, migration and anchorage independent growth, and that overexpression of the T286D phosphomimic leads to a further increase in the invasive, migratory and anchorage independent growth capacity of these cells. Pharmacological inhibition of CaMKII decreases MDA-MB-231 migration and invasion. Furthermore, we demonstrate that overexpression of T286D, but not WT or T286V-CaMKII, leads to phosphorylation of FAK, STAT5a, and Akt. These results demonstrate a novel function for phosphorylation of CaMKII at T286 in the control of breast cancer metastasis, offering a promising target for the development of therapeutics to prevent breast cancer metastasis.

No MeSH data available.


Related in: MedlinePlus

T286D phosphomimic mutation of CaMKII enhances breast cancer cell invasion.(A) MDA-MB-231 and (B) MCF-7 cells expressing CaMKII mutants were examined for ability to invade through Matrigel plugs. n = 3. *denotes statistical significance p < 0.05, **p < 0.01, ***p < 0.001, as determined by one-way ANOVA.
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f5: T286D phosphomimic mutation of CaMKII enhances breast cancer cell invasion.(A) MDA-MB-231 and (B) MCF-7 cells expressing CaMKII mutants were examined for ability to invade through Matrigel plugs. n = 3. *denotes statistical significance p < 0.05, **p < 0.01, ***p < 0.001, as determined by one-way ANOVA.

Mentions: Similarly, significantly greater numbers of MDA-MB-231 (Fig. 5A) and MCF-7 (Fig. 5B) cells overexpressing WT-CaMKII invaded through Matrigel plugs when compared to control EV cells (p < 0.05, for both), and phosphomimic mutation of T286 further enhanced invasion of both cell lines when compared to WT and T286V expressing cells (Fig. 5A,B; p < 0.001, for both). This is the first evidence identifying cellular functions controlled by pT286-CaMKII in cancer cells.


Phosphorylation of calcium/calmodulin-stimulated protein kinase II at T286 enhances invasion and migration of human breast cancer cells
T286D phosphomimic mutation of CaMKII enhances breast cancer cell invasion.(A) MDA-MB-231 and (B) MCF-7 cells expressing CaMKII mutants were examined for ability to invade through Matrigel plugs. n = 3. *denotes statistical significance p < 0.05, **p < 0.01, ***p < 0.001, as determined by one-way ANOVA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5015093&req=5

f5: T286D phosphomimic mutation of CaMKII enhances breast cancer cell invasion.(A) MDA-MB-231 and (B) MCF-7 cells expressing CaMKII mutants were examined for ability to invade through Matrigel plugs. n = 3. *denotes statistical significance p < 0.05, **p < 0.01, ***p < 0.001, as determined by one-way ANOVA.
Mentions: Similarly, significantly greater numbers of MDA-MB-231 (Fig. 5A) and MCF-7 (Fig. 5B) cells overexpressing WT-CaMKII invaded through Matrigel plugs when compared to control EV cells (p < 0.05, for both), and phosphomimic mutation of T286 further enhanced invasion of both cell lines when compared to WT and T286V expressing cells (Fig. 5A,B; p < 0.001, for both). This is the first evidence identifying cellular functions controlled by pT286-CaMKII in cancer cells.

View Article: PubMed Central - PubMed

ABSTRACT

Calcium/calmodulin-stimulated protein kinase II (CaMKII) is a multi-functional kinase that controls a range of cellular functions, including proliferation, differentiation and apoptosis. The biological properties of CaMKII are regulated by multi-site phosphorylation. However, the role that CaMKII phosphorylation plays in cancer cell metastasis has not been examined. We demonstrate herein that CaMKII expression and phosphorylation at T286 is increased in breast cancer when compared to normal breast tissue, and that increased CAMK2 mRNA is associated with poor breast cancer patient prognosis (worse overall and distant metastasis free survival). Additionally, we show that overexpression of WT, T286D and T286V forms of CaMKII in MDA-MB-231 and MCF-7 breast cancer cells increases invasion, migration and anchorage independent growth, and that overexpression of the T286D phosphomimic leads to a further increase in the invasive, migratory and anchorage independent growth capacity of these cells. Pharmacological inhibition of CaMKII decreases MDA-MB-231 migration and invasion. Furthermore, we demonstrate that overexpression of T286D, but not WT or T286V-CaMKII, leads to phosphorylation of FAK, STAT5a, and Akt. These results demonstrate a novel function for phosphorylation of CaMKII at T286 in the control of breast cancer metastasis, offering a promising target for the development of therapeutics to prevent breast cancer metastasis.

No MeSH data available.


Related in: MedlinePlus