The human neuroblastoma SK-SY5Y cell line bears functional endothelin-A-receptors and endothelin.
The study reported here characterizes the presence both of endothelin (ET) receptors and of a synthesizing ET apparatus in the human neuroblastoma SK-SY5Y cell line. We demonstrated, using reverse transcriptase polymerase chain reaction (RT-PCR), that these cells bound [125I]ET-1. The potency order of ET analogs to inhibit [125I]ET-1 binding was consistent with the presence of ET(A)-receptors. [Ca2+]i was increased by both ET-1 and ET-3 (potency order: ET-1 > ET-3. The mRNAs of preproendothelin-1 and of endothelin converting enzyme (ECE) were expressed by cells, as shown by RT-PCR studies. These mRNAs were translated into functional proteins as the cells were able to release mature (1-21) ET-like immunoreactivity into the culture medium. That secretion was time-dependent and was enhanced by treatment of the cells by the phorbol ester 12-O-tetradecanoyl phorbol-13-acetate. These results show that the human SK-SY5Y neuroblastoma cell line produces mature ET which could act as an autocrine/paracrine factor these cells.
Hamroun
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D
INSERM U.469, CCIPE, Montpellier France.
Mathieu
M N
MN
Chevillard
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C
D002118:Calcium / Q000378:metabolism; D019332:Endothelin-1 / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D009447:Neuroblastoma / Q000378:metabolism / Q000473:pathology; D044022:Receptor, Endothelin A; D017466:Receptors, Endothelin / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D019332:Endothelin-1; D044022:Receptor, Endothelin A; D017466:Receptors, Endothelin; D002118:Calcium