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Published on: August 1, 2017
Regulation of interleukin 6 expression in murine bone marrow stromal cells
J M Gimble1, J Hudson, J Henthorn
1Immunobiology and Cancer Program, Oklahoma Medical Research Foundation, Oklahoma City 73104.
Insights
Interleukin 6 (IL-6) expression in stromal cells is regulated by cytokines and chemical agents, affecting B-lymphopoiesis. This regulation involves chromatin changes and is similar to human cells.
Area of Science:
- Molecular Biology
- Immunology
- Cell Biology
Background:
- Murine stromal cell line BMS2 supports B-lymphopoiesis.
- Interleukin 6 (IL-6) plays a role in immune responses.
Purpose of the Study:
- To investigate the regulation of IL-6 expression in BMS2 cells.
- To analyze IL-6 mRNA and protein levels in response to various stimuli.
- To explore IL-6 gene regulation at the chromatin level.
Main Methods:
- Kinetic analysis of IL-6 mRNA induction and decay.
- Measurement of IL-6 biological activity.
- Cloning of the murine IL-6 genomic gene.
- Analysis of DNA nicking and topoisomerase I activity.
Main Results:
- Tumor necrosis factor, interleukin 1 (alpha and beta), and transforming growth factor beta, along with forskolin and dibutyryl cyclic AMP, transiently increase IL-6 mRNA and protein.
- IL-6 expression induction correlates with increased DNA nicking, suggesting topoisomerase I and nuclease involvement.
- Camptothecin, a topoisomerase I inhibitor, was used to support these findings.
Conclusions:
- IL-6 regulation in murine stromal cells supporting B-lymphopoiesis is comparable to human diploid fibroblasts.
- Chromatin-level regulation, including DNA nicking and topoisomerase I activity, influences IL-6 expression.
Abstract:
The regulation of interleukin 6 (IL-6) expression in the B-lymphocyte-supporting murine stromal cell line BMS2 has been examined in response to exogenous cytokines and chemical agents. Kinetic analyses of IL-6 mRNA induction and decay are presented together with analysis of the IL-6 biological activity. The cytokines tumor necrosis factor, interleukin 1 (alpha and beta), and transforming growth factor beta, as well as forskolin and dibutyryl cyclic AMP, all induce a transient rise in the steady-state level of IL-6 mRNA and an increased release of IL-6 protein. To study its regulation at the chromatin level, the murine IL-6 genomic gene has been cloned. Induction of IL-6 expression correlates with increased DNA nicking, consistent with increased topoisomerase I and endogenous nuclease activity. This finding is supported by kinetic analyses using camptothecin, a topoisomerase I inhibitor. We conclude that IL-6 regulation in murine stromal cells capable of supporting B-lymphopoiesis is comparable to that observed in human diploid fibroblasts.
