Hepatocyte growth factor disrupts cell contact and stimulates an increase in type 3 inositol triphosphate receptor expression, intracellular calcium levels, and apoptosis of rat ovarian surface epithelial cells.
The present studies revealed that hepatocyte growth factor (HGF) disrupts cell contact, increases both type 3 IP3 receptor and intracellular calcium ([Ca2+]i) levels and induces apoptosis of rat ovarian surface epithelial cells (ROSE-179 cells). Type 3 IP3 receptor was only increased in cells that lost cell contact. Disrupting cell contact by depleting extracellular calcium (Ca2+) also resulted in an increase in [Ca2+]i levels and an increase in apoptosis. These responses were prevented by the addition of 0.7 mM Ca2+. Actinomycin D and cycloheximide prevented apoptosis that resulted from Ca2+ removal. In situ hybridization studies revealed that type 3 IP3 receptor was expressed at relatively low levels by ROSE-179 cells cultured with Ca2+ but at high levels in the absence of Ca2+. ROSE-179 cells cultured in Ca2+-free medium with type 3 IP3 receptor antisense oligonucleotide lost cell contact but did not show an increase in either type 3 IP3 receptor protein, [Ca2+]i, or apoptosis. The nonsense oligonucleotide did not alter these responses to Ca2+ removal. Thus, the disruption of cell contact by either HGF or Ca2+ depletion increases the expression of type 3 IP3 receptor, which causes an increase in [Ca2+]i and the apoptotic death of ROSE-179 cells.
Lail-Trecker
M R
MR
Department of Physiology, University of Connecticut Health Center, Farmington, CT, USA.
Peluso
C E
CE
Peluso
J J
JJ
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002118:Calcium / Q000008:administration & dosage / Q000378:metabolism*; D015220:Calcium Channels / Q000235:genetics*; D002448:Cell Adhesion / Q000187:drug effects*; D002460:Cell Line; D003470:Culture Media; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D017403:In Situ Hybridization; D053496:Inositol 1,4,5-Trisphosphate Receptors; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010053:Ovary / Q000166:cytology* / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D015220:Calcium Channels; D003470:Culture Media; D053496:Inositol 1,4,5-Trisphosphate Receptors; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear; D003609:Dactinomycin; D017228:Hepatocyte Growth Factor; D003513:Cycloheximide; D002118:Calcium