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Interaction Between Mevalonate Pathway and Retinoic Acid-Induced Differentiation.

Gueddari-Pouzols N, Duriez P, Chomienne C, Trussardi A, Jardillier JC - J. Biomed. Biotechnol. (2001)

Bottom Line: The supply of an excess of cholesterol (up to 80 &mgr;g/ml of LDL) leads to a significant decrease of cell differentiation by ATRA from 78% +/-0.1 to 54% +/-2.8.A concomitant decrease of cell growth (51% +/- 6.4) was observed.The pretreatment of cells by the geranylgeranyltransferase inhibitor (GGTI-298) has no effect on the cell differentiation process.

View Article: PubMed Central - HTML - PubMed

ABSTRACT
All trans retinoic acid (ATRA) is a potent inducer of differentiation of HL-60 cell line. The pretreatment of the cells by compactin, a competitive inhibitor of 3-hydroxy-3-methylglutaryl (HMG) CoA reductase, during 24 hours, enhances the ATRA-induced cell differentiation. At 50 nM, the percentage of cell differentiation is 34.9% +/- 2 and 73% +/- 2.96 in the control and compactin-treated cells, respectively. The removal of compactin boosts the level of HMG-CoA reductase and therefore the biosynthesis of sterol and nonsterol isoprenoid compounds. The participation of sterol and nonsterol pathway was then investigated. The supply of an excess of cholesterol (up to 80 &mgr;g/ml of LDL) leads to a significant decrease of cell differentiation by ATRA from 78% +/-0.1 to 54% +/-2.8. A concomitant decrease of cell growth (51% +/- 6.4) was observed. The pretreatment of cells by the geranylgeranyltransferase inhibitor (GGTI-298) has no effect on the cell differentiation process. By contrast, the farnesyltransferase inhibitors (FTI-II and FTI-277) completely abolish theATRA-induced differentiation, thus confirming the involvement of farnesylated proteins in the differentiation mechanism.

No MeSH data available.


Effect of ATRA on HL-60 cell differentiation. HL-60 cells were incubated for 24 hours in foetal calf lipoprotein deprived serum (LDS medium) supplemented with compactin (2 μM). Then, they were washed in phosphate buffered saline and cultured during 5 days in LDS medium containing increasing concentrations of ATRA (▴). The control cells were cultured during 5 days in FCS medium containing increasing concentrations of ATRA (Δ). At day 5, cell differentiation was determined by NBT reduction and viable cell number as determined by Trypan blue exclusion. Insert: Effect on cell growth. ATRA-treated cells 50 nM (▪) and control cells (□).
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Figure 1: Effect of ATRA on HL-60 cell differentiation. HL-60 cells were incubated for 24 hours in foetal calf lipoprotein deprived serum (LDS medium) supplemented with compactin (2 μM). Then, they were washed in phosphate buffered saline and cultured during 5 days in LDS medium containing increasing concentrations of ATRA (▴). The control cells were cultured during 5 days in FCS medium containing increasing concentrations of ATRA (Δ). At day 5, cell differentiation was determined by NBT reduction and viable cell number as determined by Trypan blue exclusion. Insert: Effect on cell growth. ATRA-treated cells 50 nM (▪) and control cells (□).

Mentions: The presence of compactin and cholesterol-deprived medium (LDS) had no effect on the basal differentiation activity of HL-60 cells. The treatment of HL-60 cells with ATRA induced a gradual increase of differentiated cells. When endogenouscholesterol synthesis was inhibited, in a first step for 24 hours by incubation in LDS containing compactin (2 μM), then the removal of compactin induced an increaseof the sterol and nonsterol pathways, a possible consequence of the overexpression of HMG-CoA reductase, ATRA induced cell differentiation but the effect was more pronounced in the cells temporarily treated with compactin. At 50 nM, ATRA induced, respectively, 34.9% ± 2 and 73% ± 2.96 of cell differentiation (Figure 1), in the control and compactin-treated cells, whereas 1000 nM was required to obtain 80% ± 4 in control cells.


Interaction Between Mevalonate Pathway and Retinoic Acid-Induced Differentiation.

Gueddari-Pouzols N, Duriez P, Chomienne C, Trussardi A, Jardillier JC - J. Biomed. Biotechnol. (2001)

Effect of ATRA on HL-60 cell differentiation. HL-60 cells were incubated for 24 hours in foetal calf lipoprotein deprived serum (LDS medium) supplemented with compactin (2 μM). Then, they were washed in phosphate buffered saline and cultured during 5 days in LDS medium containing increasing concentrations of ATRA (▴). The control cells were cultured during 5 days in FCS medium containing increasing concentrations of ATRA (Δ). At day 5, cell differentiation was determined by NBT reduction and viable cell number as determined by Trypan blue exclusion. Insert: Effect on cell growth. ATRA-treated cells 50 nM (▪) and control cells (□).
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC129055&req=5

Figure 1: Effect of ATRA on HL-60 cell differentiation. HL-60 cells were incubated for 24 hours in foetal calf lipoprotein deprived serum (LDS medium) supplemented with compactin (2 μM). Then, they were washed in phosphate buffered saline and cultured during 5 days in LDS medium containing increasing concentrations of ATRA (▴). The control cells were cultured during 5 days in FCS medium containing increasing concentrations of ATRA (Δ). At day 5, cell differentiation was determined by NBT reduction and viable cell number as determined by Trypan blue exclusion. Insert: Effect on cell growth. ATRA-treated cells 50 nM (▪) and control cells (□).
Mentions: The presence of compactin and cholesterol-deprived medium (LDS) had no effect on the basal differentiation activity of HL-60 cells. The treatment of HL-60 cells with ATRA induced a gradual increase of differentiated cells. When endogenouscholesterol synthesis was inhibited, in a first step for 24 hours by incubation in LDS containing compactin (2 μM), then the removal of compactin induced an increaseof the sterol and nonsterol pathways, a possible consequence of the overexpression of HMG-CoA reductase, ATRA induced cell differentiation but the effect was more pronounced in the cells temporarily treated with compactin. At 50 nM, ATRA induced, respectively, 34.9% ± 2 and 73% ± 2.96 of cell differentiation (Figure 1), in the control and compactin-treated cells, whereas 1000 nM was required to obtain 80% ± 4 in control cells.

Bottom Line: The supply of an excess of cholesterol (up to 80 &mgr;g/ml of LDL) leads to a significant decrease of cell differentiation by ATRA from 78% +/-0.1 to 54% +/-2.8.A concomitant decrease of cell growth (51% +/- 6.4) was observed.The pretreatment of cells by the geranylgeranyltransferase inhibitor (GGTI-298) has no effect on the cell differentiation process.

View Article: PubMed Central - HTML - PubMed

ABSTRACT
All trans retinoic acid (ATRA) is a potent inducer of differentiation of HL-60 cell line. The pretreatment of the cells by compactin, a competitive inhibitor of 3-hydroxy-3-methylglutaryl (HMG) CoA reductase, during 24 hours, enhances the ATRA-induced cell differentiation. At 50 nM, the percentage of cell differentiation is 34.9% +/- 2 and 73% +/- 2.96 in the control and compactin-treated cells, respectively. The removal of compactin boosts the level of HMG-CoA reductase and therefore the biosynthesis of sterol and nonsterol isoprenoid compounds. The participation of sterol and nonsterol pathway was then investigated. The supply of an excess of cholesterol (up to 80 &mgr;g/ml of LDL) leads to a significant decrease of cell differentiation by ATRA from 78% +/-0.1 to 54% +/-2.8. A concomitant decrease of cell growth (51% +/- 6.4) was observed. The pretreatment of cells by the geranylgeranyltransferase inhibitor (GGTI-298) has no effect on the cell differentiation process. By contrast, the farnesyltransferase inhibitors (FTI-II and FTI-277) completely abolish theATRA-induced differentiation, thus confirming the involvement of farnesylated proteins in the differentiation mechanism.

No MeSH data available.