Ghrelin is produced by the human erythroleukemic HEL cell line and involved in an autocrine pathway leading to cell proliferation.
Ghrelin, a ligand of the GH secretagogue receptor (GHS-R 1a), is a 28-amino acid peptide with an unusual octanoyl group on Ser3, crucial for its biological activity. For the first time, ghrelin and GHS-R 1b, a truncated variant of the receptor resulting from alternative splicing, but not GHS-R 1a, mRNAs were detected in the human erythroleukemic cell line HEL. Two antibodies, used for RIA, were directed against octanoylated and total (octanoylated and desoctanoylated) ghrelin, and the recognized epitopes were characterized. Using reverse phase HPLC analysis followed by RIA, we demonstrated that octanoylated and desoctanoylated ghrelins were present in HEL cells and their culture medium, of which more than 90% was octanoylated. The ghrelin levels were not affected after 24 h treatment with sodium butyrate, phorbol 12-myristate 13-acetate, or forskolin, but a significant 3-fold increase in desoctanoylated ghrelin was detected in the culture medium after 48 h treatment with sodium butyrate. The antighrelin SB801 and SB969 antisera inhibited HEL cell proliferation by 24% and 39%, respectively, after 72 h. Taken together, these data suggested that endogenous ghrelin stimulated HEL cell proliferation by an autocrine pathway involving an unidentified receptor, distinct from GHS-R1a, and that the HEL cell line represents a unique model to study the octanoylation of ghrelin.
De Vriese
Carine
C
Department of Biochemistry and Nutrition, Faculty of Medicine, Université Libre de Bruxelles, Bat G/E, CP 611, 808 route de Lennik, B-1070 Brussels, Belgium.
Grégoire
Françoise
F
De Neef
Philippe
P
Robberecht
Patrick
P
Delporte
Christine
C
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000906:Antibodies / Q000737:chemistry; D001667:Binding, Competitive; D002091:Butyrylcholinesterase / Q000378:metabolism; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D054439:Ghrelin; D006801:Humans; D007700:Kinetics; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism*; D008969:Molecular Sequence Data; D036361:Peptide Hormones / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D011863:Radioimmunoassay; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012978:Sodium Oxybate / Q000737:chemistry; D013997:Time Factors
D000906:Antibodies; D003470:Culture Media; D054439:Ghrelin; D036361:Peptide Hormones; D010455:Peptides; D012333:RNA, Messenger; D012978:Sodium Oxybate; D002265:Carboxylic Ester Hydrolases; D002091:Butyrylcholinesterase