Research report| Volume 117, ISSUE 1, P125-138, September 1995

Download started.


1,25-Dihydroxyvitamin D3-induced HL-60 macrophages: regulation of cholesterol and LDL metabolism

  • Zeinab E. Jouni
    Department of Nutritional Sciences and Interdisciplinary Nutritional Sciences Program, 309 Shantz Building, The University of Arizona, Tucson, AZ 85721, USA
    Search for articles by this author
  • Joy J. Winzerling
    Department of Nutritional Sciences and Interdisciplinary Nutritional Sciences Program, 309 Shantz Building, The University of Arizona, Tucson, AZ 85721, USA
    Search for articles by this author
  • Donald J. McNamara
    Corresponding author. Tel.: +1 602 621 1971; Fax: +1 602 621 9446.
    Department of Nutritional Sciences and Interdisciplinary Nutritional Sciences Program, 309 Shantz Building, The University of Arizona, Tucson, AZ 85721, USA
    Search for articles by this author
  • Author Footnotes
    1 Current address: Department of Biochemistry, University of Arizona, Tucson, AZ 85721, USA.
      This paper is only available as a PDF. To read, Please Download here.


      Differentiation of human promyelocytic leukemic HL-60 cells with 1,25-dihydroxyvitamin D3 (D3) results in macrophages which exhibit specific and saturable receptor-mediated processing of both native and modified low density lipoprotein (LDL). Analysis of binding kinetics demonstrated that macrophages bind LDL and acetyl-LDL with similar affinities, yet possess significantly different numbers of receptors (55 ± 6 × 103 LDL receptors/cell vs. 79 ± 7 × 103 acetyl-LDL receptors/cell). D3-induced HL-60 macrophages challenged with LDL or acetyl-LDL exhibited suppression of HMG-CoA reductase activity as well as a significant induction in the incorporation of [14C]oleate into cholesteryl ester compared with macrophages incubated with lipoprotein depleted serum. Maximum increases in ACAT activity were obtained in macrophages incubated with 25-hydroxycholesterol plus LDL or acetyl-LDL. The increase in ACAT activity in macrophages challenged with acetyl-LDL paralleled the increase in cellular cholesterol content and the increase of oil red O lipid stainable material, imparting the macrophages with a foamy appearance. The data indicate that D3-induced HL-60 macrophages are a useful model for the study of lipoprotein -macrophage interactions as related to foam cell development and atherogenesis.


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Atherosclerosis
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Harberland ME
        • Fogelman AM
        The role of altered lipoproteins in the pathogenesis of atherosclerosis.
        Am Heart J. 1987; 113: 573
        • Fogelman AM
        • Harberland ME
        • Seager J
        • Hokom M
        • Edwards PA
        Factors regulating the activities of low density lipoprotein receptor and the scavenger receptor on human monocyte-macrophages.
        J Lipid Res. 1981; 22: 1131
        • Steinberg D
        • Parathasarathy S
        • Carew TE
        • Khoo JC
        • Witztum JL
        Modifications of low-density lipoprotein that increase atherogenicity.
        N Engl J Med. 1989; 320: 915
        • Ellsworth JL
        • Cooper AD
        • Kraemer FB
        Evidence that chylomicron remnants and B-VLDL are transported by the same receptor pathway in J774 murine macrophagederived cells.
        J Lipid Res. 1986; 27: 1062
        • Brown MS
        • Goldstein JL
        Lipoprotein metabolism in the macrophage. Implications for cholesterol deposition in atherosclerosis.
        Ann Rev Biochem. 1983; 52: 223
        • Jouni ZE
        • McNamara DJ
        Lipoprotein receptors of HL-60 macrophages: effect of differentiation with tetramyristic phorbol acetate and 1,25-dihydroxyvitamin D3.
        Arteriosclerosis Thromb. 1991; 11: 995
        • Goldstein JL
        • Basu SK
        • Brown MS
        Receptor-mediated endocytosis of low density lipoprotein in culture cell.
        Methods Enzymol. 1983; 98: 241
        • Brown MS
        • Goldstein JL
        • Krieger M
        • Ho YK
        • Anderson RGW
        Reversible accumulation of cholesteryl esters in macrophages incubated with acetylated lipoproteins.
        J Cell Biol. 1979; 82: 597
        • Brown MS
        • Ho YK
        • Goldstein JL
        The cholesteryl ester cycle in macrophage foam cells.
        J Biol Chem. 1980; 255: 9344
        • Luskey KL
        • Faust JR
        • Chin DJ
        • Brown MS
        • Goldstein JL
        Amplification of the gene for 3-hydroxy-3-methylglutaryl coenzyme A reductase, but not for 53-KDa protein, in UT-1 cells.
        J Biol Chem. 1983; 258: 8462
        • Gil G
        • Faust JR
        • Chin DJ
        • Goldstein JL
        • Brown MS
        Membrane-bound domain of HMG-CoA reductase is required for sterol-enhanced degradation of the enzyme.
        Cell. 1985; 41: 249
        • Russel DW
        • Yamamoto T
        • Schneider WJ
        • Slaughter CJ
        • Brown MS
        • Goldstein JL
        cDNA cloning of the bovine low density lipoprotein receptor: Feedback regulation of a receptor mRNA.
        in: Proc Natl Acad Sci USA. 80. 1983: 7501
        • Goldstein JL
        • Ho YK
        • Basu SK
        • Brown MS
        Binding site on macrophages that mediates uptake and degradation of acetylated low density lipoprotein, producing massive cholesterol deposition.
        in: Proc Natl Acad Sci USA. 76. 1979: 333
        • Mangelsdorf DJ
        • Koffler HP
        • Donaldson CA
        • Pike JW
        • Haussler MR
        1,25-Dihydroxyvitamin D3-induced differentiation in a human promyelocytic leukemia cell line (HL-60): receptor-mediated maturation to macrophage-like cells.
        J Cell Biol. 1984; 98: 391
        • Havel RJ
        • Eder HA
        • Bragdon JH
        The distribution and chemical composition of ultracentrifugally separated lipoproteins in human serum.
        J Clin Invest. 1955; 34: 1345
        • Mills GL
        • Lane PA
        • Weech PK
        The isolation and purification of plasma lipoproteins.
        in: Laboratory Techniques in Biochemistry and Molecular Biology. A Guidebook to Lipoprotein Technique. Elsevier, New York1984: 55
        • Basu SK
        • Goldstein JL
        • Anderson RG
        • Brown MS
        Degradation of cationized low density lipoprotein and regulation of cholesterol metabolism in homozygous familial hypercholesterolemia fibroblasts.
        in: Proc Natl Acad Sci USA. 73. 1976: 3178
        • Keightley DD
        • Fisher RJ
        • Cressie NAC
        Properties and interpretation of the Woolf and Scatchard plots in analyzng data from steroid receptor assays.
        J Steroid Biochem. 1983; 19: 1407
        • Yachnin S
        • Toub DB
        • Mannickarottu V
        Divergence in cholesterol biosynthesis rates and 3(OH)-HMG-CoA reductase activity as a consequence of granulocyte versus monocyte-macrophage differentiation in HL-60 cells.
        in: Proc Natl Acad Sci USA. 81. 1984: 889
        • Shapiro DJ
        • Nordstrom JL
        • Mitschelen JJ
        • Rodwell VW
        • Schimke RT
        Micro assay for 3-hydroxy-3-methylglutaryl CoA reductase in rat liver and in L-cell fibroblasts.
        Biochim Biophys Acta. 1974; 370: 369
        • Chautan M
        • Termine E
        • Nalbone G
        • LaFont H
        Acyl coenzyme A:cholesterol acyl transferase assay: silica gel column separation of reaction products.
        Anal Biochem. 1988; 173: 436
        • McNamara DJ
        • Ahrens Jr, EH
        • Morrissey K
        Role of the kidney in the metabolism of plasma mevalonate. Studies in human and rhesus monkeys.
        J Clin Invest. 1985; 76: 31
        • Markwell MAK
        • Haas SM
        • Bieber LL
        • Tolbert NE
        A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples.
        Anal Biochem. 1978; 87: 206
        • Glantz SA
        Primer of Biostatistics.
        McGraw-Hill, New York1981
        • Patel DD
        • Knight BL
        The effect of mevalonate on 3-hydroxy-3-methylglutaryl-CoA reductase activity and the absolute rate of cholesterol biosynthesis in human monocyte-derived macrophage.
        Eur J Biochem. 1985; 153: 117
        • Knight BL
        • Patel DD
        • Soutar AK
        The regulation of 3-hydroxy-3-methylglutaryl-CoA reductase activity, cholesterol esterification and the expression of low-density lipoprotein receptors in cultures of monocyte-derived macrophages.
        Biochem J. 1983; 210: 523
        • Tsukada T
        • Rsenfeld M
        • Ross R
        • Gown AM
        Immunocytochemical analysis of cellular components in lesions. Use of monoclonal antibodies with the Watanable and fat-fed rabbit.
        Arteriosclerosis. 1986; 6: 601
        • Hashimoto S
        • Dayton S
        • Alfin-Slater RB
        • Bui PT
        • Baker N
        • Wilson L
        Characteristics of the cholesterol-esterifying activity in normal and atherosclerotic rabbit aortas.
        Circ Res. 1974; 34: 176
        • Hasimoto S
        • Drevon CA
        • Weinstein DB
        • Bernett JS
        • Dayton S
        • Steinberg D
        Activity of acyl-CoA:cholesterol acyltransferase and 3-hydroxy-3-methylglutaryl-CoA reductase in subfractions of hepatic microsomes enriched with cholesterol.
        Biochim Biophys Acta. 1983; 754: 126
        • Chang TY
        • Limanek JS
        • Chang CCY
        Evidence indicating that inactivation of 3-hydroxy-3-methylglutaryl coenzyme A reductase by low density lipoproteins or by 25-hydroxycholesterol requires mediator protein(s) with rapid turnover rate.
        J Biol Chem. 1981; 256: 6174
        • Suckling KE
        • Boyd GS
        • Smellie CG
        Properties of a solubilized and reconstituted preparation of acyl-CoA:cholesterol acyltransferase from rat liver.
        Biochim Biophys Acta. 1982; 710: 154
        • Doolittle GM
        • Chan TY
        Solubilization, partial purification, and reconstitution in phosphatidyl-cholesterol liposomes of acyl-CoA:cholesterol acyltransferase.
        Biochemistry. 1982; 21: 674
        • Roullet JB
        • Haluska M
        • Morchoisne O
        • McCarron DA
        1,25-Dihydroxyvitamin D3-induced alterations of lipid metabolism in human monocyte-macrophages.
        Am J Physiol. 1989; 25: E290
        • Miyaura C
        • Abe E
        • Momoi T
        • Suda T
        Alteration of lipid metabolism associated with the activation of mouse alveolar macrophages induced by 1a,25-dihydroxyvitamin D3.
        Endocrinology. 1987; 120: 1813
        • Hara H
        • Tanishita H
        • Yokoyama S
        • Tajima S
        • Yamamoto A
        Induction of acetylated low density lipoprotein receptor on the cells of human monocytic leukemia cell line (THP-1 cell).
        Biochem Biophys Res Commun. 1987; 146: 802
        • Goldstein JL
        • Brown MS
        Stanbury JB Wyngaarden JB Fredrickson DS Goldstein JL Brown MS The metabolic basis of inherited disease. McGraw-Hill, New York1983: 672
        • Auwrex JH
        • Chait A
        • Deeb SS
        Regulation of the low density lipoprotein receptor and hydroxymethylglutaryl coenzyme A reductase gene by protein kinase C and putative regulatory protein.
        in: Proc Natl Acad Sci USA. 861. 1989: 1133
        • Via PD
        • Plant AL
        • Craig IF
        • Gotto AM
        • Smith LC
        Metabolism of normal and modified low density lipoproteins by macrophage cell lines of murine and human origin.
        Biochim Biophys Acta. 1985; 833: 417
        • Hayashi K
        • Dojo S
        • Hirata Y
        • Ohtani H
        • Nakashima K
        • Nishio E
        • Kurushima H
        • Saeki M
        • Kajiyama G
        Metabolic changes in LDL receptors and an appearance of scavenger receptors after phorbol ester-induced differentiation of U937 cells.
        Biochim Biophys Acta. 1991; 1082: 152