p60(v-src) and serum control cell shape and apoptosis via distinct pathways in quail neuroretina cells.
We made use of QNR cells transformed by a thermosensitive (tsNY68) strain of the Rous sarcoma virus (RSV) to compare the effect of p60(v-src) and serum in cultured nerve cells. In this system, both p60(v-src) heat inactivation and serum removal resulted in growth arrest in G1. In both cases, growth arrest was reversible since cell proliferation was rapidly re-induced following respectively p60v-src renaturation or serum re-addition. However, cells did not fully recover their ability to grow in soft agar, suggesting that, in contrast to the cell cycle machinery, the transforming capacities of these cells have been irreversibly altered. We found that p60(v-src) kinase activity prevented detachment from the substratum and cell death following serum removal. Thermal inactivation of p60(v-src) at restrictive temperature (41.5 degrees C), but not serum removal, resulted in dramatic morphological changes, which occurred 4 h after temperature shift up to 41.5 degrees C. Later on, typical features of apoptotic cells could be observed. Cell death was greatly reduced by the caspase-3 inhibitor ZVAD.FMK, but not by the caspase-1 inhibitor Ac-YVAD.CHO. Together, these results suggested that p60(v-src) and serum factors act on distinct pathways, at least in part. In an attempt to identify the signalling pathways involved in the cell response to p60(v-src) down regulation, we found that Erk and Rac were rapidly inactivated following temperature shift up to 41.5 degrees C. Thus, the combined effects of p60(v-src) and serum factors on the cytoskeleton dynamics and the apoptosis machinery are essential for full neoplastic transformation of neuroretina cells.
Aouacheria
Abdel
A
Institut de Biologie et Chimie des Protéines, UMR 5086 CNRS-Université Claude Bernard 7, passage du Vercors F69367 Lyon cedex 07, France.
Ory
Stéphane
S
Schmitt
Jean-Robert
JR
Rigal
Dominique
D
Jurdic
Pierre
P
Gillet
Germain
G
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001358:Avian Sarcoma Viruses / Q000235:genetics; D001798:Blood Proteins / Q000494:pharmacology*; D015153:Blotting, Western; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D048429:Cell Size / Q000187:drug effects; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000473:pathology; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D015688:Oncogene Protein pp60(v-src) / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011489:Protein Denaturation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt; D011784:Quail; D012160:Retina / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D015398:Signal Transduction / Q000187:drug effects; D013696:Temperature; D013997:Time Factors
D001798:Blood Proteins; D061945:Caspase Inhibitors; D016895:Culture Media, Serum-Free; D015688:Oncogene Protein pp60(v-src); D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D020928:Mitogen-Activated Protein Kinases; D053148:Caspase 3; D020169:Caspases