PLC-gamma1 is required for IGF-I protection from cell death induced by loss of extracellular matrix adhesion.
Phospholipase C-gamma1, a tyrosine kinase substrate, hydrolyses phosphatidylinositol 4,5-bisphosphate to produce inositol 1,4,5-trisphosphate and diacylglycerol, which act as second messenger moleculesto mobilize intracellular calcium and activate protein kinase C, respectively. We have investigated the role of phospholipase C-gamma1 in anoikis, or cell death, induced by the loss of extracellular matrix adhesion. Spontaneously immortalized mouse embryonic fibroblasts nullizygous at the Plcg1 locus (Plcg1(-/-)), referred to as Null cells, were derived from targeted gene disruption experiments. Subsequently, phospholipase C-gamma1 was re-expressed in these cells to derive Null+ cells. The Null and Null+ cells were then placed in suspension to induce cell death, which was measured directly as well as by the induction of caspase 3, as an index of programmed cell death or apoptosis. The results demonstrate that insulin-like growth factor can rescue Null+ cells but not Null cells from suspension-induced cell death. This demonstrates that phospholipase C-gamma1 is required for insulin-like growth factor dependent cell survival under these conditions. Lastly, the data demonstrate that insulinlike growth factor stimulated tyrosine phosphorylation of phospholipase C-gamma1 in both adherent and suspension cells.
Chattopadhyay
Ansuman
A
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-0146, USA.
Carpenter
Graham
G
D000818:Animals; D023102:Anoikis; D002448:Cell Adhesion; D002460:Cell Line; D002470:Cell Survival; D021982:Cell-Matrix Junctions; D005109:Extracellular Matrix / Q000201:enzymology / Q000378:metabolism*; D005347:Fibroblasts; D017353:Gene Deletion; D007334:Insulin-Like Growth Factor I / Q000378:metabolism*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D020935:MAP Kinase Signaling System; D051379:Mice; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D051966:Phospholipase C gamma; D010738:Type C Phospholipases / Q000235:genetics / Q000378:metabolism*
D007527:Isoenzymes; D007334:Insulin-Like Growth Factor I; D019869:Phosphatidylinositol 3-Kinases; D010738:Type C Phospholipases; D051966:Phospholipase C gamma