Downregulation of internal enhancer activity contributes to abnormally low immunoglobulin expression in the MedB-1 mediastinal B-cell lymphoma cell line.
Primary mediastinal B-cell lymphoma (PMBL) is a highly aggressive tumour with a unique pattern of clinical, morphological, immunological and genetic features distinct from other diffuse large B-cell lymphomas. PMBLs are characterized by a mature B-cell phenotype, but they typically lack immunoglobulin (Ig) gene expression. The PMBL cell line MedB-1 shares many characteristic properties of the primary tumour, including low-level Ig production despite a functionally rearranged IgVH gene and absence of 'crippling' mutations. In this study, a search was undertaken for reasons for downregulated Ig expression. Similar levels of the B-cell-specific transcription factors BOB.1/OBF.1 and PU.1 were found in MedB-1 cells to those in the Ig-producing UM-1 lymphoblastoid cell line. However, MedB-1 lacked the Oct2 transcription factor. Reporter assays showed that Ig-type promoters were active in MedB-1 cells. In contrast, activity of the intronic heavy chain enhancer was dramatically reduced. Ectopic expression of Oct2 was able partially to restore enhancer activity but transcription from the endogenous IgVH gene could not be rescued. Therefore, the role of epigenetic factors in the downregulation of Ig was investigated. Methylated histone 3 lysine 9, a reliable marker of chromatin silencing, was not detected in MedB-1 promoter and enhancer regions. Inhibition of DNA methyltransferase and of histone deacetylases also did not reactivate Ig production. These data suggest the existence of alternative mechanisms of Ig inhibition in MedB-1 cells, different from chromatin silencing and the lack of Oct2.
Ritz
Olga
O
Department of Physiological Chemistry, University of Ulm, Germany.
Leithäuser
Frank
F
Hasel
Cornelia
C
Brüderlein
Silke
S
Ushmorov
Alexey
A
Möller
Peter
P
Wirth
Thomas
T
D019175:DNA Methylation; D015536:Down-Regulation / Q000276:immunology; D004742:Enhancer Elements, Genetic; D006801:Humans; D007136:Immunoglobulins / Q000096:biosynthesis* / Q000235:genetics; D016393:Lymphoma, B-Cell / Q000235:genetics* / Q000276:immunology; D008479:Mediastinal Neoplasms / Q000235:genetics* / Q000276:immunology; D009363:Neoplasm Proteins / Q000378:metabolism; D050813:Octamer Transcription Factor-2 / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014157:Transcription Factors / Q000378:metabolism; D014407:Tumor Cells, Cultured
D007136:Immunoglobulins; D009363:Neoplasm Proteins; D050813:Octamer Transcription Factor-2; D014157:Transcription Factors