Hormone sensitive lipase mRNA in both monocyte and macrophage forms of the human THP-1 cell line

William J. Johnson, So Young Jang, David W. Bernard

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

The identity of the neutral cholesteryl ester hydrolase (CEH) in human monocyte/macrophages is uncertain. Prior studies indicate that hormone sensitive lipase (HSL) is a major CEH in mouse macrophages, and that HSL mRNA is present in human THP-1 monocytes. In the present study, HSL mRNA expression was examined in THP-1 cells as a function of differentiation status and cholesterol enrichment. By RT-PCR with primer pairs that span exon boundaries, HSL mRNA was demonstrated in THP-1 monocytes and phorbol-ester differentiated THP-1 macrophages. cDNA identities were confirmed by sequencing. By Northern blotting, with HSL cDNA as probe, THP-1 monocytes were found to contain HSL mRNA of approximately 3 and 3.9 kb. In THP-1 macrophages, the 3 kb mRNA was greatly diminished, while the level of the 3.9 kb mRNA was maintained. mRNA of approximately 3 and 3.9 kb are those expected of the 86-kDa (adipocyte) and 117-kDa (testicular) HSL isoforms, respectively. The presence of the testicular isoform mRNA was confirmed in THP-1 cells by amplification and sequencing of an isoform-specific cDNA. Additionally, Northern-blot comparisons showed that the 3 and 3.9 kb mRNA in THP-1 comigrated with the HSL mRNA in 3T3-L1 adipocytes and rat testis, respectively. The level of the 3.9 kb mRNA did not vary greatly with cholesterol enrichment. Thus, the HSL gene is transcribed in THP-1 cells both before and after differentiation into macrophages; after differentiation, the predominant mRNA is that for the 117-kDa isoform. This isoform is a CEH, and may mediate some CE turnover in THP-1 cells. (C) 2000 Elsevier Science Inc.

Original languageEnglish (US)
Pages (from-to)543-552
Number of pages10
JournalComparative Biochemistry and Physiology - B Biochemistry and Molecular Biology
Volume126
Issue number4
DOIs
StatePublished - Jan 1 2000

Keywords

  • 3T3-L1
  • Atherosclerosis
  • Cholesteryl ester hydrolase
  • Hormone sensitive lipase
  • J774
  • Macrophage
  • Monocyte
  • mRNA
  • Testis
  • THP-1

ASJC Scopus subject areas

  • Biochemistry
  • Physiology

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