Effect of diethylstilbestrol (DES) on intracellular Ca(2+) levels in renal tubular cells.
The effect of the estrogen diethylstilbestrol (DES) on intracellular Ca(2+) concentrations ([Ca(2+)](i)) in Madin Darby canine kidney (MDCK) cells was investigated, using the fluorescent dye fura-2 as a Ca(2+) indicator. DES (10-50 microM) evoked [Ca(2+)](i) increases in a concentration-dependent manner. Extracellular Ca(2+) removal inhibited 45 +/- 5% of the Ca(2+) response. In Ca(2+)-free medium, pretreatment with 50 microM DES abolished the [Ca(2+)](i) increases induced by 2 microM carbonylcyanide m-chlorophenylhydrazone (CCCP; a mitochondrial uncoupler) and 1 microM thapsigargin (an endoplasmic reticulum Ca(2+) pump inhibitor); and pretreatment with CCCP and thapsigargin partly inhibited DES-induced [Ca(2+)](i) signals. Adding 3 mM Ca(2+) increased [Ca(2+)](i) in cells pretreated with 50 microM DES in Ca(2+)-free medium, suggesting that DES may induce capacitative Ca(2+) entry. 17beta-Estradiol (2-20 microM) increased [Ca(2+)](i), but 100 microM diethylstilbestrol dipropionate had no effect. Pretreatment with the phospholipase C inhibitor U73122 (1 microM) to abolish inositol 1,4,5-trisphosphate formation inhibited 30% of DES-induced Ca(2+) release. DES (20 microM) also increased [Ca(2+)](i) in human normal hepatocytes and osteosarcoma cells. Cumulatively, this study shows that DES induced rapid and sustained [Ca(2+)](i) increases by releasing intracellular Ca(2+) and triggering extracellular Ca(2+) entry in renal tubular cells.
Jan
C R
CR
Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Taiwan. crjan@isca.vghks.gov.tw
Cheng
J S
JS
Roan
C J
CJ
Lee
K C
KC
Chen
W C
WC
Chou
K J
KJ
Tang
K Y
KY
Wang
J L
JL
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D004054:Diethylstilbestrol / Q000494:pharmacology*; D004285:Dogs; D004958:Estradiol / Q000494:pharmacology; D005456:Fluorescent Dyes; D016257:Fura-2; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D007684:Kidney Tubules / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism
D005456:Fluorescent Dyes; D004958:Estradiol; D004054:Diethylstilbestrol; D015544:Inositol 1,4,5-Trisphosphate; D002118:Calcium; D016257:Fura-2