Identification of LIL-STAT in monocytic leukemia cells and monocytes after stimulation with interleukin-6 or
1Division of Hematology, Department of Internal Medicine, Academic Hospital Groningen, The Netherlands.
Abstract:
In acute myelogenous leukemia (AML) and adult T-cell leukemia, it has been demonstrated that the transcription factor LIL-STAT is constitutively activated. To identify and characterize this unknown LIL-STAT protein, electrophoretic mobility shift assay (EMSA) and oligoprecipitation assays were performed by using lipopolysaccharide/interleukin-1 (IL-1)-responsive element (LILRE) oligonucleotide probes. EMSA demonstrated a significant increase in LIL-STAT binding to the LILRE oligonucleotides after interferon gamma (IFN-gamma) and IL-6 stimulation of THP-1 cells. In unstimulated THP-1 and AML cells, LILRE oligonucleotide probes bound only to STAT1 alpha and beta isoforms. The LILRE element showed a significant increase in binding of both alpha and beta isoforms of STAT1 and STAT3 upon IFN-gamma and IL-6 stimulation. Similar results were observed with human monocytes upon IL-6 or IFN-gamma stimulation. These studies indicate that LIL-STAT consists of STAT1 and STAT3 proteins that bind to the LILRE DNA consensus site in a stimulus-dependent way.
