title: PIPKIγ regulates focal adhesion dynamics and colon cancer cell invasion.
abstract: Focal adhesion assembly and disassembly are essential for cell migration and cancer invasion, but the detailed molecular mechanisms regulating these processes remain to be elucidated. Phosphatidylinositol phosphate kinase type Iγ (PIPKIγ) binds talin and is required for focal adhesion formation in EGF-stimulated cells, but its role in regulating focal adhesion dynamics and cancer invasion is poorly understood. We show here that overexpression of PIPKIγ promoted focal adhesion formation, whereas cells expressing either PIPKIγ(K188,200R) or PIPKIγ(D316K), two kinase-dead mutants, had much fewer focal adhesions than those expressing WT PIPKIγ in CHO-K1 cells and HCT116 colon cancer cells. Furthermore, overexpression of PIPKIγ, but not PIPKIγ(K188,200R), resulted in an increase in both focal adhesion assembly and disassembly rates. Depletion of PIPKIγ by using shRNA strongly inhibited formation of focal adhesions in HCT116 cells. Overexpression of PIPKIγ(K188,200R) or depletion of PIPKIγ reduced the strength of HCT116 cell adhesion to fibronection and inhibited the invasive capacities of HCT116 cells. PIPKIγ depletion reduced PIP₂ levels to ∼40% of control and PIP₃ to undetectable levels, and inhibited vinculin localizing to focal adhesions. Taken together, PIPKIγ positively regulates focal adhesion dynamics and cancer invasion, most probably through PIP₂-mediated vinculin activation.
authors.affiliation: Markey Cancer Center, University of Kentucky, Lexington, Kentucky, United States of America.
authors.forename: Zhaofei
authors.initials: Z
authors.lastname: Wu
authors.forename: Xiang
authors.initials: X
authors.lastname: Li
authors.forename: Manjula
authors.initials: M
authors.lastname: Sunkara
authors.forename: Heather
authors.initials: H
authors.lastname: Spearman
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Morris
authors.forename: Cai
authors.initials: C
authors.lastname: Huang
mesh_terms: D000818:Animals; D016466:CHO Cells; D002465:Cell Movement / Q000235:genetics / Q000502:physiology; D003110:Colonic Neoplasms / Q000235:genetics / Q000378:metabolism*; D006224:Cricetinae; D005353:Fibronectins / Q000378:metabolism; D022001:Focal Adhesions / Q000235:genetics / Q000378:metabolism*; D045325:HCT116 Cells; D006801:Humans; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000378:metabolism*; D016596:Vinculin / Q000378:metabolism
chemical_list: D005353:Fibronectins; D019269:Phosphatidylinositol 4,5-Diphosphate; D016596:Vinculin; D017853:Phosphotransferases (Alcohol Group Acceptor)