1alpha,25-dihydroxyvitamin D3 induces sphingomyelin hydrolysis in HaCaT cells via tumor necrosis factor alpha.
Treatment of the human keratinocyte cell line HaCaT with 1alpha, 25-dihydroxyvitamin D3 (1,25-(OH)2D3) resulted in the hydrolysis of sphingomyelin with peak elevations of ceramide levels after 2-3 h (Geilen, C. C., Bektas, M., Wieder, Th., and Orfanos, C. E. (1996) FEBS Lett. 378, 88-92). In the present paper, the mechanism underlying the effect of 1,25-(OH)2D3 on sphingomyelin hydrolysis was investigated. Using the cell culture supernatant of HaCaT cells treated with 1,25-(OH)2D3 for 2 h, it was possible to induce sphingomyelin hydrolysis as early as 30-60 min after addition to resting cells. Several lines of experimental evidence indicated that tumor necrosis factor alpha (TNFalpha) mediates sphingomyelin hydrolysis after 1,25-(OH)2D3 treatment: (i) 1,25-(OH)2D3 stimulated TNFalpha mRNA expression after 1 h, (ii) newly synthesized TNFalpha occurred 2 h after 1,25-(OH)2D3 treatment, (iii) indirect activation of sphingomyelin hydrolysis by the supernatant of 1, 25-(OH)2D3-treated HaCaT cells was abolished by preincubation of the supernatant with antibodies directed against TNFalpha, and (iv) preincubation of HaCaT cells with neutralizing antibodies directed against the 55-kDa receptor of TNFalpha blocked the ability of 1, 25-(OH)2D3 to induce sphingomyelin hydrolysis in HaCaT cells. These data demonstrate that 1,25-(OH)2D3 activated sphingomyelin hydrolysis by an autocrine mechanism via TNFalpha expression. Furthermore, this indirect mode of action may serve as an explanation for the delayed induction of sphingomyelin hydrolysis by vitamin D3.
Geilen
C C
CC
Department of Dermatology, University Medical Center Benjamin Franklin, The Free University of Berlin, D-12200 Berlin, Germany.
Bektas
M
M
Wieder
T
T
Kodelja
V
V
Goerdt
S
S
Orfanos
C E
CE
D002117:Calcitriol / Q000494:pharmacology*; D002460:Cell Line; D003545:Cysteine / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D012333:RNA, Messenger; D013109:Sphingomyelins / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
D012333:RNA, Messenger; D013109:Sphingomyelins; D014409:Tumor Necrosis Factor-alpha; D008715:Methionine; D002117:Calcitriol; D003545:Cysteine