20-Hydroxycholecalciferol, Product of Vitamin D3 Hydroxylation by P450scc, Decreases NF-kB Activity by Increasing IkBa Levels in Human Keratinocytes

Z. Janjetovic, M.A. Zmijewski, Robert Tuckey, D.A. Deleon, Minh Nguyen, L.M. Pfeffer, A.T. Slominski

Research output: Contribution to journalArticle

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Abstract

The side chain of vitamin D3 is hydroxylated in a sequential manner by cytochrome P450scc (CYP11A1) to form 20-hydroxycholecalciferol, which can induce growth arrest and differentiation of both primary and immortalized epidermalkeratinocytes. Since nuclear factor-kB (NF-kB) plays a pivotal role in the regulation of cell proliferation, differentiation andapoptosis, we examined the capability of 20-hydroxycholecalciferol to modulate the activity of NF-kB, using 1,25-dihydroxycholecalciferol (calcitriol) as a positive control. 20-hydroxycholecalciferol inhibits the activation of NFkB DNAbinding activity as well as NF-kB-driven reporter gene activity in keratinocytes. Also, 20-hydroxycholecalciferol inducedsignificant increases in the mRNA and protein levels of the NF-kB inhibitor protein, IkBa, in a time dependent manner, whileno changes in total NF-kB-p65 mRNA or protein levels were observed. Another measure of NF-kB activity, p65 translocationfrom the cytoplasm into the nucleus was also inhibited in extracts of 20-hydroxycholecalciferol treated keratinocytes.Increased IkBa was concomitantly observed in cytosolic extracts of 20-hydroxycholecalciferol treated keratinocytes, asdetermined by immunoblotting and immunofluorescent staining. In keratinocytes lacking vitamin D receptor (VDR), 20-hydroxycholecalciferol did not affect IkBa mRNA levels, indicating that it requires VDR for its action on NF-kB activity.Comparison of the effects of calcitrol, hormonally active form of vitamin D3, with 20-hydrocholecalciferol show that bothagents have a similar potency in inhibiting NF-kB. Since NF-kB is a major transcription factor for the induction ofinflammatory mediators, our findings indicate that 20-hydroxycholecalciferol may be an effective therapeutic agent forinflammatory and hyperproliferative skin diseases.
Original languageEnglish
Pages (from-to)Article number e5988, 12pp
JournalP L o S One
Volume4
Issue number6
DOIs
Publication statusPublished - 2009

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hydroxycholecalciferols
Hydroxylation
Hydroxycholecalciferols
cholecalciferol
Cholecalciferol
keratinocytes
hydroxylation
Keratinocytes
Cholesterol Side-Chain Cleavage Enzyme
calcitriol
Calcitriol Receptors
Calcitriol
vitamin D
Messenger RNA
receptors
Proteins
proteins
extracts
Cell proliferation
skin diseases

Cite this

Janjetovic, Z. ; Zmijewski, M.A. ; Tuckey, Robert ; Deleon, D.A. ; Nguyen, Minh ; Pfeffer, L.M. ; Slominski, A.T. / 20-Hydroxycholecalciferol, Product of Vitamin D3 Hydroxylation by P450scc, Decreases NF-kB Activity by Increasing IkBa Levels in Human Keratinocytes. In: P L o S One. 2009 ; Vol. 4, No. 6. pp. Article number e5988, 12pp.
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abstract = "The side chain of vitamin D3 is hydroxylated in a sequential manner by cytochrome P450scc (CYP11A1) to form 20-hydroxycholecalciferol, which can induce growth arrest and differentiation of both primary and immortalized epidermalkeratinocytes. Since nuclear factor-kB (NF-kB) plays a pivotal role in the regulation of cell proliferation, differentiation andapoptosis, we examined the capability of 20-hydroxycholecalciferol to modulate the activity of NF-kB, using 1,25-dihydroxycholecalciferol (calcitriol) as a positive control. 20-hydroxycholecalciferol inhibits the activation of NFkB DNAbinding activity as well as NF-kB-driven reporter gene activity in keratinocytes. Also, 20-hydroxycholecalciferol inducedsignificant increases in the mRNA and protein levels of the NF-kB inhibitor protein, IkBa, in a time dependent manner, whileno changes in total NF-kB-p65 mRNA or protein levels were observed. Another measure of NF-kB activity, p65 translocationfrom the cytoplasm into the nucleus was also inhibited in extracts of 20-hydroxycholecalciferol treated keratinocytes.Increased IkBa was concomitantly observed in cytosolic extracts of 20-hydroxycholecalciferol treated keratinocytes, asdetermined by immunoblotting and immunofluorescent staining. In keratinocytes lacking vitamin D receptor (VDR), 20-hydroxycholecalciferol did not affect IkBa mRNA levels, indicating that it requires VDR for its action on NF-kB activity.Comparison of the effects of calcitrol, hormonally active form of vitamin D3, with 20-hydrocholecalciferol show that bothagents have a similar potency in inhibiting NF-kB. Since NF-kB is a major transcription factor for the induction ofinflammatory mediators, our findings indicate that 20-hydroxycholecalciferol may be an effective therapeutic agent forinflammatory and hyperproliferative skin diseases.",
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20-Hydroxycholecalciferol, Product of Vitamin D3 Hydroxylation by P450scc, Decreases NF-kB Activity by Increasing IkBa Levels in Human Keratinocytes. / Janjetovic, Z.; Zmijewski, M.A.; Tuckey, Robert; Deleon, D.A.; Nguyen, Minh; Pfeffer, L.M.; Slominski, A.T.

In: P L o S One, Vol. 4, No. 6, 2009, p. Article number e5988, 12pp.

Research output: Contribution to journalArticle

TY - JOUR

T1 - 20-Hydroxycholecalciferol, Product of Vitamin D3 Hydroxylation by P450scc, Decreases NF-kB Activity by Increasing IkBa Levels in Human Keratinocytes

AU - Janjetovic, Z.

AU - Zmijewski, M.A.

AU - Tuckey, Robert

AU - Deleon, D.A.

AU - Nguyen, Minh

AU - Pfeffer, L.M.

AU - Slominski, A.T.

PY - 2009

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N2 - The side chain of vitamin D3 is hydroxylated in a sequential manner by cytochrome P450scc (CYP11A1) to form 20-hydroxycholecalciferol, which can induce growth arrest and differentiation of both primary and immortalized epidermalkeratinocytes. Since nuclear factor-kB (NF-kB) plays a pivotal role in the regulation of cell proliferation, differentiation andapoptosis, we examined the capability of 20-hydroxycholecalciferol to modulate the activity of NF-kB, using 1,25-dihydroxycholecalciferol (calcitriol) as a positive control. 20-hydroxycholecalciferol inhibits the activation of NFkB DNAbinding activity as well as NF-kB-driven reporter gene activity in keratinocytes. Also, 20-hydroxycholecalciferol inducedsignificant increases in the mRNA and protein levels of the NF-kB inhibitor protein, IkBa, in a time dependent manner, whileno changes in total NF-kB-p65 mRNA or protein levels were observed. Another measure of NF-kB activity, p65 translocationfrom the cytoplasm into the nucleus was also inhibited in extracts of 20-hydroxycholecalciferol treated keratinocytes.Increased IkBa was concomitantly observed in cytosolic extracts of 20-hydroxycholecalciferol treated keratinocytes, asdetermined by immunoblotting and immunofluorescent staining. In keratinocytes lacking vitamin D receptor (VDR), 20-hydroxycholecalciferol did not affect IkBa mRNA levels, indicating that it requires VDR for its action on NF-kB activity.Comparison of the effects of calcitrol, hormonally active form of vitamin D3, with 20-hydrocholecalciferol show that bothagents have a similar potency in inhibiting NF-kB. Since NF-kB is a major transcription factor for the induction ofinflammatory mediators, our findings indicate that 20-hydroxycholecalciferol may be an effective therapeutic agent forinflammatory and hyperproliferative skin diseases.

AB - The side chain of vitamin D3 is hydroxylated in a sequential manner by cytochrome P450scc (CYP11A1) to form 20-hydroxycholecalciferol, which can induce growth arrest and differentiation of both primary and immortalized epidermalkeratinocytes. Since nuclear factor-kB (NF-kB) plays a pivotal role in the regulation of cell proliferation, differentiation andapoptosis, we examined the capability of 20-hydroxycholecalciferol to modulate the activity of NF-kB, using 1,25-dihydroxycholecalciferol (calcitriol) as a positive control. 20-hydroxycholecalciferol inhibits the activation of NFkB DNAbinding activity as well as NF-kB-driven reporter gene activity in keratinocytes. Also, 20-hydroxycholecalciferol inducedsignificant increases in the mRNA and protein levels of the NF-kB inhibitor protein, IkBa, in a time dependent manner, whileno changes in total NF-kB-p65 mRNA or protein levels were observed. Another measure of NF-kB activity, p65 translocationfrom the cytoplasm into the nucleus was also inhibited in extracts of 20-hydroxycholecalciferol treated keratinocytes.Increased IkBa was concomitantly observed in cytosolic extracts of 20-hydroxycholecalciferol treated keratinocytes, asdetermined by immunoblotting and immunofluorescent staining. In keratinocytes lacking vitamin D receptor (VDR), 20-hydroxycholecalciferol did not affect IkBa mRNA levels, indicating that it requires VDR for its action on NF-kB activity.Comparison of the effects of calcitrol, hormonally active form of vitamin D3, with 20-hydrocholecalciferol show that bothagents have a similar potency in inhibiting NF-kB. Since NF-kB is a major transcription factor for the induction ofinflammatory mediators, our findings indicate that 20-hydroxycholecalciferol may be an effective therapeutic agent forinflammatory and hyperproliferative skin diseases.

U2 - 10.1371/journal.pone.0005988

DO - 10.1371/journal.pone.0005988

M3 - Article

VL - 4

SP - Article number e5988, 12pp

JO - P L o S One

JF - P L o S One

SN - 1932-6203

IS - 6

ER -