The spontaneous expression of interleukin-1 beta and interleukin-6 is associated with spontaneous expression of AP-1 and NF-kappa B transcription factor in acute myeloblastic leukemia cells.
The nature of the spontaneous expression of cytokines that is observed in blasts of some AML patients is unclear. We studied whether or not the spontaneous expression of IL-1 beta and IL-6 is due to an increased transcription rate of the cytokine gene and associated with a spontaneous expression of two transcription factors that play an important role in IL-1 beta and IL-6 gene transcription, namely activator protein-1 (AP-1) and nuclear factor-kappa B (NF-kappa B). In eight of the 19 AML patients a spontaneous expression of IL-1 beta mRNA was observed, whereas IL-6 mRNA was expressed in seven of the cases. Expression of IL-6 mRNA correlated nicely with the secretion of IL-6 protein. Nuclear run-on experiments showed that spontaneous expression of IL-1 beta and IL-6 was at least partly due to an increased transcription rate of the respective genes compared to the results from healthy unstimulated monocytes. Electrophoretic mobility shift assays demonstrated that especially spontaneous expression of NF-kappa B is associated with spontaneous cytokine expression. However, the spontaneous expression of transcription factors is not due to the endogenous secretion of IL-1 since the addition of anti-IL-1 monoclonal antibody did not affect the expression of NF-kappa B. Finally, supershift experiments were performed that demonstrated that the NF-kappa B consists of the p50 and the p65 subunits. In summary, these results demonstrate that the spontaneous expression of cytokines is frequently associated with an increased transcription rate and a spontaneous expression of transcription factors.
Dokter
W H
WH
Department of Medicine, University of Groningen, The Netherlands.
Tuyt
L
L
Sierdsema
S J
SJ
Esselink
M T
MT
Vellenga
E
E
D000208:Acute Disease; D001483:Base Sequence; D015973:Gene Expression Regulation, Leukemic; D017781:Genes, Immediate-Early; D006801:Humans; D007375:Interleukin-1 / Q000096:biosynthesis* / Q000235:genetics; D015850:Interleukin-6 / Q000096:biosynthesis* / Q000235:genetics; D007951:Leukemia, Myeloid / Q000473:pathology*; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics; D014411:Neoplastic Stem Cells / Q000378:metabolism*; D011759:Pyrrolidines / Q000494:pharmacology; D013859:Thiocarbamates / Q000494:pharmacology; D018808:Transcription Factor AP-1 / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D014158:Transcription, Genetic / Q000187:drug effects; D014407:Tumor Cells, Cultured
D007375:Interleukin-1; D015850:Interleukin-6; D016328:NF-kappa B; D009363:Neoplasm Proteins; D011759:Pyrrolidines; D013859:Thiocarbamates; D018808:Transcription Factor AP-1; C020972:pyrrolidine dithiocarbamic acid