Diverse expression of G-proteins in human sarcoma cell lines with different osteogenic potential: evidence for the involvement of Gi2 in cell proliferation.
Previously, it has been shown that the GTP-binding protein Gi2 is implicated in cellular growth [1,2] and differentiation [2,3]. In the present paper we demonstrate that this is also the case for human sarcoma cells. Six human osteosarcoma and three soft tissue sarcoma clonal cell lines were analyzed for levels of G-protein mRNA and polypeptide expression and effector enzyme (i.e., adenylate cyclase and phospholipase C) activation, which were all compared with individual growth rates. Unexpectedly, it appeared that the various strains exhibited large inter-individual variations in G-protein expression and signaling system activation. However, cell doubling time in the exponential phase of growth was inversely correlated (r = 0.71, P < 0.05) to immunodetected levels of intrinsic Gi2 alpha. Furthermore, cells stably transfected with a retroviral (pZipNeo(SV)X) construct containing the activating or inactivating Gi2 alpha-R179E or Gi2 alpha-G204A point mutations consistently reduced or enhanced individual cell strain doubling time, respectively. It appeared that other parameters investigated, including cellular alkaline phosphatase and monoclonal antibody epitope binding, both being markers of the proliferating osteoblast, did not correlate with cell doubling times.
Gordeladze
J O
JO
Institute of Medical Biochemistry, University of Oslo, Norway.
Lund
H W
HW
Jablonski
G
G
Bruland
O S
OS
D000262:Adenylyl Cyclases / Q000378:metabolism; D001483:Base Sequence; D001859:Bone Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D002455:Cell Division / Q000235:genetics; D019204:GTP-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D012516:Osteosarcoma / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D012983:Soft Tissue Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D014407:Tumor Cells, Cultured; D010738:Type C Phospholipases / Q000378:metabolism
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D010738:Type C Phospholipases; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases