RRC ID 49230
Author Ueno N, Nishimura N, Ueno S, Endo S, Tatetsu H, Hirata S, Hata H, Matsuoka M, Mitsuya H, Okuno Y.
Title PU.1 acts as tumor suppressor for myeloma cells through direct transcriptional repression of IRF4.
Journal Oncogene
Abstract We previously reported that PU.1 is downregulated in the majority of myeloma cell lines and primary myeloma cells of certain myeloma patients, and conditional expression of PU.1 in such myeloma cell lines induced cell cycle arrest and apoptosis. We found downregulation of IRF4 protein in the U266 myeloma cell line following induction of PU.1. Previous studies reported that knockdown of IRF4 in myeloma cell lines induces apoptosis, prompting us to further investigate the role of IRF4 downregulation in PU.1-induced cell cycle arrest and apoptosis in myeloma cells. PU.1 induced downregulation of IRF4 at the protein level, cell cycle arrest and apoptosis in six myeloma cell lines. Chromatin immunoprecipitation (ChIP) revealed that PU.1 directly binds to the IRF4 promoter, whereas a reporter assay showed that PU.1 may suppress IRF4 promoter activity. Stable expression of IRF4 in myeloma cells expressing PU.1 partially rescued the cells from apoptosis induced by PU.1. As it was reported that IRF4 directly binds to the IRF7 promoter and downregulates its expression in activated B cell-like subtype of diffuse large B cell lymphoma cells, we performed ChIP assays and found that IRF4 directly binds the IRF7 promoter in myeloma cells. It is known that IRF7 positively upregulates interferon-β (IFNβ) and induces apoptosis in many cell types. Binding of IRF4 to the IRF7 promoter decreased following PU.1 induction, accompanied by downregulation of IRF4 protein expression. Knockdown of IRF7 protected PU.1-expressing myeloma cells from apoptosis. Furthermore, IFNβ, which is a downstream target of IRF7, was upregulated in myeloma cells along with IRF7 after PU.1 induction. Finally, we evaluated the mRNA expression levels of PU.1, IRF4 and IRF7 in primary myeloma cells from patients and found that PU.1 and IRF7 were strongly downregulated in contrast to the high expression levels of IRF4. These data strongly suggest that PU.1-induced apoptosis in myeloma cells is associated with IRF4 downregulation and subsequent IRF7 upregulation.
Volume 36(31)
Pages 4481-4497
Published 2017-8-1
DOI 10.1038/onc.2017.79
PII onc201779
PMID 28368411
MeSH Apoptosis Humans Interferon Regulatory Factor-7 / genetics Interferon Regulatory Factors / genetics Interferon Regulatory Factors / physiology* Interferon-beta / biosynthesis Multiple Myeloma / genetics* Multiple Myeloma / pathology Promoter Regions, Genetic Proto-Oncogene Proteins / physiology* Trans-Activators / physiology* Transcription, Genetic Tumor Suppressor Proteins / physiology* U937 Cells
IF 7.971
Times Cited 8
DNA material CSII-EF-MCS-IRES2-Venus (RDB04384) pCMV-VSV-G (RDB04392) pCAG-HIVgp (RDB04394).