Studies of protein kinase C alpha as a potential target for inhibition of murine melanoma metastasis.
Item
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Title
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Studies of protein kinase C alpha as a potential target for inhibition of murine melanoma metastasis.
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Identifier
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AAI3144142
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identifier
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3144142
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Creator
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Sullivan, Regina M.
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Contributor
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Adviser: Susan A. Rotenberg
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Date
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2004
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Language
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English
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Publisher
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City University of New York.
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Subject
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Biology, Cell
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Abstract
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Pharmacological and genetic techniques were used to establish the role of PKCalpha in the migration and adhesion of murine melanoma cells (B16F10). PKC is a monomeric serine, threonine protein kinase that exists as a family of isoforms. An isoform profile revealed PKCalpha as both the most abundantly expressed isoform and the only conventional isoform. The pharmacological approach to these studies involved the use of the PKC inhibitor, dequalinium, which previously was shown to inhibit the catalytic activity of highly purified PKC. Using analogues to the parent compound C 10-DECA, PKCalpha is shown to be the major intracellular target of the most potent analogue C14-DECA. At a concentration of 250nM and in combination with long-wave UV light, which causes irreversible binding of the inhibitor, inactivation of intracellular PKCalpha is correlated to inhibition of migration. C14-DECA is also shown to inhibit the metastasis of B16F10 cells in C57BL/6 mice. In a genetic approach, B16F10 cells were stably transfected to overexpress a kinase-defective PKCalpha. In migration assays these cells show reduced migration on both collagen IV and fibronectin. Furthermore, these transfections show reduced phosphorylation of focal adhesion kinase (FAK). The phosphorylation of FAK is likely to be an autocatalytic event that is critical to the formation of focal adhesions. Taken together, these results define PKCalpha as a mechanistic component in the migration pathway of murine melanoma cells and implicate FAK as a downstream effector of PKCalpha in this pathway.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
