Inhibition of integrin-linked kinase/protein kinase B/Akt signaling: mechanism for ganglioside-induced apoptosis.
Ganglioside GT1b inhibits keratinocyte attachment to and migration on a fibronectin matrix by binding to alpha(5)beta(1) and preventing alpha(5)beta(1) interaction with fibronectin. The role of gangliosides in triggering keratinocyte apoptosis, however, is unknown. Addition of GT1b to keratinocyte-derived SCC12 cells, grown in serum-free medium but exposed to fibronectin, suppressed Bad phosphorylation, activated caspase-9, and inhibited cyclin D and E expression, resulting in cell cycle arrest at G(1) phase and initiation of apoptosis. The mechanism of GT1b activation of caspase-9 involved inhibition of beta(1) integrin serine/threonine phosphorylation and decreased phosphorylation of both integrin-linked kinase and protein kinase B/Akt at its Ser-473 site, leading to cytochrome c release from mitochondria. Consistently, blockade of GT1b function with anti-GT1b antibody specifically activated the Ser-473 site of Akt, markedly suppressing apoptosis. The ganglioside-induced inhibition of Akt phosphorylation was GT1b-specific and was not observed when cells were treated with other keratinocyte gangliosides, including GD3. These studies suggest that the modulation of keratinocyte cell cycle and survival by GT1b is mediated by its direct interaction with alpha(5)beta(1) and resultant inhibition of the integrin/integrin-linked kinase/protein kinase B/Akt signaling pathway.
Wang
X Q
XQ
Department of Pediatrics, Children's Memorial Institute for Education and Research, Northwestern University Medical School, Chicago, Illinois 60614, USA.
Sun
P
P
Paller
A S
AS
D017209:Apoptosis; D001665:Binding Sites; D053453:Caspase 9; D020169:Caspases / Q000378:metabolism; D002453:Cell Cycle; D002460:Cell Line; D002465:Cell Movement; D002470:Cell Survival; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D056741:Cyclin D; D019927:Cyclin E / Q000378:metabolism; D016213:Cyclins / Q000378:metabolism; D003574:Cytochrome c Group / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004792:Enzyme Precursors / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D016193:G1 Phase; D005732:Gangliosides / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D015603:Keratinocytes / Q000378:metabolism; D010766:Phosphorylation; D011233:Precipitin Tests; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt; D012694:Serine / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction; D013912:Threonine / Q000737:chemistry / Q000378:metabolism; D013997:Time Factors; D000099200:Scaffold Protein ILK
D053453:Caspase 9; D020169:Caspases; D016895:Culture Media, Serum-Free; D056741:Cyclin D; D019927:Cyclin E; D016213:Cyclins; D003574:Cytochrome c Group; D004792:Enzyme Precursors; D005353:Fibronectins; D005732:Gangliosides; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D051057:Proto-Oncogene Proteins c-akt; D012694:Serine; D013912:Threonine; D000099200:Scaffold Protein ILK; C494918:AKT1 protein, human; C506433:CASP9 protein, human