U18666A inhibits intracellular cholesterol transport and neurotransmitter release in human neuroblastoma cells.
To determine if neurochemical function might be impaired in cell models with altered cholesterol balance, we studied the effects of U18666A (3-beta-[(2-diethyl-amino)ethoxy]androst-5-en-17-one) on intracellular cholesterol metabolism in three human neuroblastoma cell lines (SK-N-SH, SK-N-MC, and SH-SY5Y). U18666A (< or =0.2 microg/ml) completely inhibited low density lipoprotein (LDL)-stimulated cholesterol esterification in SK-N-SH cells, while cholesterol esterification stimulated by 25-hydroxycholesterol or bacterial sphingomyelinase was unaffected or partially inhibited, respectively. U18666A also blocked LDL-stimulated downregulation of LDL receptor and caused lysosomal accumulation of cholesterol as measured by filipin staining. U18666A treatment for 18 h resulted in 70% inhibition of K+-evoked norepinephrine release in phorbol ester-differentiated SH-SY5Y cells, while release stimulated by the calcium ionophore A23187 was only slightly affected. These results suggest that U 18666A may preferentially block a voltage-regulated Ca2+ channel involved in norepinephrine release and that alterations in neurotransmitter secretion might be a feature of disorders such as Niemann-Pick Type C, in which intracellular cholesterol transport and distribution are impaired.
Sparrow
S M
SM
Atlantic Research Centre, Department of Pediatrics, Dalhousie University, Halifax, Nova Scotia, Canada.
Carter
J M
JM
Ridgway
N D
ND
Cook
H W
HW
Byers
D M
DM
D000736:Androstenes / Q000494:pharmacology*; D000924:Anticholesteremic Agents / Q000494:pharmacology*; D001692:Biological Transport / Q000187:drug effects; D000001:Calcimycin / Q000494:pharmacology; D015220:Calcium Channels / Q000502:physiology; D002784:Cholesterol / Q000378:metabolism*; D003470:Culture Media; D015536:Down-Regulation; D006801:Humans; D006888:Hydroxycholesterols / Q000494:pharmacology; D007700:Kinetics; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism; D009447:Neuroblastoma / Q000378:metabolism*; D009638:Norepinephrine / Q000378:metabolism*; D011188:Potassium / Q000494:pharmacology; D011973:Receptors, LDL / Q000187:drug effects / Q000378:metabolism*; D013108:Sphingomyelin Phosphodiesterase / Q000494:pharmacology; D013211:Staphylococcus aureus / Q000201:enzymology; D014407:Tumor Cells, Cultured