在内毒素和干扰素-γ的作用下，集落刺激因子-1衍生的巨噬细胞在分化过程中对原癌基因和细胞因子基因的转录水平进行差异调节。

Endotoxin and interferon-gamma differentially regulate the transcriptional levels of proto-oncogenes and cytokine genes during the differentiation of colony-stimulating factor type-1-derived macrophages.

作者信息

Myers M J, Ghildyal N, Schook L B

机构信息

Department of Animal Sciences, University of Illinois, Urbana, USA.

出版信息

Immunology. 1995 Jun;85(2):318-24.

PMID:7642223

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1383898/

Abstract

The expression of cytokine genes for interleukin-1 (IL-1) (alpha and beta) and tumour necrosis factor-alpha (TNF-alpha), along with the proto-oncogenes c-fos, c-fms and c-myc, was examined by nuclear run-off and Northern blot analysis during in vitro differentiation of colony-stimulating factor type-1 (CSF-1)-derived bone marrow macrophages (BMDM). Constitutive transcription of c-myc was maximal on day 3 and decreased with differentiation. Constitutive transcription of c-fms and c-fos was similar at all times examined. In contrast, the steady-state mRNA levels were maximal on day 5 for c-myc and day 7 for c-fms and c-fos. Thirty minutes after endotoxin (lipopolysaccharide; LPS) stimulation, there was a rapid increase in run-off transcription rates for c-myc in day 3-day 9 BMDM, with maximal levels observed in day 7 BMDM. c-fms transcription was maximally induced within 1 hr by LPS in day 3 and day 5 BMDM. LPS induced transcription of c-fos to equivalent levels in day 3-day 9 BMDM. LPS stimulation augmented steady-state mRNA levels for c-myc, c-fms and c-fos. Maximal induction of c-myc was observed in day 3 BMDM. c-fos and c-fms were both maximally induced in day 5 and day 7 BMDM. Interferon-gamma (IFN-gamma) had no effect on transcription of the proto-oncogenes examined. In contrast to the proto-oncogenes, peak levels of run-off transcription for IL-1 alpha and IL-1 beta genes were observed 1-2 hr after LPS stimulation for day 3, day 5 and day 7 BMDM. The kinetics of LPS-induced steady-state mRNA accumulation of IL-1 alpha and IL-1 beta were similar to the kinetics of run-off transcription. Constitutive transcription of TNF-alpha was observed on all days of differentiation. LPS and IFN-gamma both enhanced run-off transcription of the TNF-alpha gene; however, LPS had a more pronounced effect. The kinetics of induction of TNF-alpha transcription paralleled the kinetics of steady-state TNF-alpha mRNA accumulation. IFN-gamma resulted in secretion of TNF-alpha in day 5, day 7, and day 9 BMDM after 4-8 hr of stimulation. Day 3 BMDM had little, if any, secreted TNF-alpha activity.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

在集落刺激因子-1（CSF-1）诱导的骨髓巨噬细胞（BMDM）体外分化过程中，通过核转录延伸分析和Northern印迹分析，检测了白细胞介素-1（IL-1）（α和β）、肿瘤坏死因子-α（TNF-α）以及原癌基因c-fos、c-fms和c-myc的细胞因子基因表达。c-myc的组成型转录在第3天达到最大值，并随着分化而降低。在所有检测时间点，c-fms和c-fos的组成型转录相似。相比之下，c-myc的稳态mRNA水平在第5天达到最大值，c-fms和c-fos的稳态mRNA水平在第7天达到最大值。内毒素（脂多糖；LPS）刺激30分钟后，第3天至第9天的BMDM中c-myc的转录延伸率迅速增加，在第7天的BMDM中观察到最高水平。在第3天和第5天的BMDM中，LPS在1小时内最大程度地诱导了c-fms转录。LPS在第3天至第9天的BMDM中将c-fos转录诱导至同等水平。LPS刺激增加了c-myc、c-fms和c-fos的稳态mRNA水平。在第3天的BMDM中观察到c-myc的最大诱导。c-fos和c-fms在第5天和第7天的BMDM中均被最大程度地诱导。干扰素-γ（IFN-γ）对所检测的原癌基因转录没有影响。与原癌基因不同，在第3天、第5天和第7天的BMDM中，LPS刺激后1 - 2小时观察到IL-1α和IL-1β基因的转录延伸峰值水平。LPS诱导的IL-1α和IL-1β稳态mRNA积累动力学与转录延伸动力学相似。在分化的所有天数均观察到TNF-α的组成型转录。LPS和IFN-γ均增强了TNF-α基因的转录延伸；然而，LPS的作用更显著。TNF-α转录的诱导动力学与稳态TNF-α mRNA积累动力学平行。IFN-γ在刺激4 - 8小时后导致第5天、第7天和第9天的BMDM分泌TNF-α。第3天的BMDM几乎没有（如果有的话）分泌的TNF-α活性。（摘要截断于400字）