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Tumor necrosis factor alpha (TNF-α) is a potent cytokine involved in systemic inflammation and immune modulation. Signaling responses that involve TNF-α are context dependent and capable of stimulating pathways promoting both cell death and survival. TNF-α treatment has been investigated as part of a combined therapy for acute myeloid leukemia due to its modifying effects on all-trans retinoic acid (ATRA) mediated differentiation into granulocytes. To investigate the interaction between cellular differentiation and TNF-α, we performed RNA-sequencing on two forms of the human HL-60/S4 promyelocytic leukemia cell line treated with TNF-α. The ATRA-differentiated granulocytic form of HL-60/S4 cells had an enhanced transcriptional response to TNF-α treatment compared to the undifferentiated promyelocytes. The observed TNF-α responses included differential expression of cell cycle gene sets, which were generally upregulated in TNF-α treated promyelocytes, and downregulated in TNF-α treated granulocytes. This is consistent with TNF-α induced cell cycle repression in granulocytes and cell cycle progression in promyelocytes. Moreover, we found evidence that TNF-α treatment of granulocytes shifts the transcriptome toward that of a macrophage. We conclude that TNF-α treatment promotes a divergent transcriptional program in promyelocytes and granulocytes. TNF-α promotes cell cycle associated gene expression in promyelocytes. In contrast, TNF-α stimulated granulocytes have reduced cell cycle gene expression, and a macrophage-like transcriptional program.


Originally published:
Elsie C. Jacobson, Lekha Jain, Mark H. Vickers, Ada L. Olins, Donald E. Olins, Jo K. Perry, Justin M. O’Sullivan (2019), TNF-α Differentially Regulates Cell Cycle Genes in Promyelocytic and Granulocytic HL-60/S4 Cells. G3: GENES, GENOMES, GENETICS August 1, 2019 vol. 9 no. 8 2775-2786;

Copyright © 2019 Jacobson et al.This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary file retrieved from:
Jacobson, Elsie C.; Jain, Lekha; Vickers, Mark H.; Olins, Ada L.; Olins, Donald E.; Perry, Jo K.; et al. (2019): Supplemental Material for Jacobsen et al., 2019. GSA Journals. Dataset.

Supplemental materials descriptions:
Supplementary tables 1-22. 1. Significantly differentially expressed genes in promyelocytes treated with TNF-a, differentiated into granulocytes and macrophages, and granulocytes treated with TNF-a. 2. RT-qPCR and RNA-seq of CDK1, TNF, and VCAM1. 3-21. Functional analysis summary tables of GSEA, GO, KEGG, and hallmark terms enriched in subsets of differentially expressed genes. 22. Primers and probes used for RT-qPCR validation.
Supplementary figure 1. PCA of VST-normalized transcript counts show conditions cluster together in A) promyelocytes treated with TNF-a, B) granulocytes treated with TNF-a, and C) promyelocytes differentiated into granulocytes and macrophages. (8986 kB)
Supplementary materials .zip file



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