TGF-beta induces apoptosis in human B cells by transcriptional regulation of BIK and BCL-XL

Cell Death Differ. 2009 Apr;16(4):593-602. doi: 10.1038/cdd.2008.183. Epub 2009 Jan 9.

Abstract

Transforming growth factor-beta (TGF-beta) potently induces apoptosis in Burkitt's lymphoma (BL) cell lines and in explanted primary human B lymphocytes. The physiological relevance and mechanism of TGF-beta-mediated apoptosis induction in these cells remains to be determined. Here we demonstrate the requirement for TGF-beta-mediated regulation of BIK and BCL-X(L) to activate an intrinsic apoptotic pathway in centroblastic BL cells. TGF-beta directly induced transcription of BIK and a consensus Smad-binding element identified in the BIK promoter recruits TGF-beta-activated Smad transcription factor complexes in vivo. TGF-beta also transcriptionally repressed expression of the apoptosis inhibitor BCL-X(L). Inhibition of BCL-X(L) sensitised BL cells to TGF-beta-induced apoptosis whereas overexpression of BCL-X(L) or suppression of BIK by shRNA, diminished TGF-beta-induced apoptosis. BIK and BCL-X(L) were also identified as TGF-beta target genes in purified normal human centroblast B cells and immunohistochemical analyses of tonsil tissue revealed widespread TGF-beta receptor-regulated Smad activation and a focal pattern of BIK expression. Furthermore, using a selective inhibitor of the TGF-beta receptor we provide evidence that autocrine TGF-beta signalling through ALK5 contributes to the default apoptotic programme in normal human centroblasts undergoing spontaneous apoptosis. Our data suggests that TGF-beta may act as a physiological mediator of human germinal centre homoeostasis by regulation of BIK and BCL-X(L).

MeSH terms

  • Apoptosis / drug effects*
  • Apoptosis Regulatory Proteins / genetics
  • Apoptosis Regulatory Proteins / metabolism*
  • B-Lymphocytes / cytology
  • B-Lymphocytes / drug effects*
  • B-Lymphocytes / metabolism
  • Cell Line, Tumor
  • Cells, Cultured
  • Electrophoretic Mobility Shift Assay
  • Flow Cytometry
  • Gene Expression Regulation / drug effects*
  • Humans
  • Immunoblotting
  • Immunohistochemistry
  • In Vitro Techniques
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Mitochondrial Proteins
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transforming Growth Factor beta / pharmacology*
  • bcl-X Protein / genetics
  • bcl-X Protein / metabolism*

Substances

  • Apoptosis Regulatory Proteins
  • BIK protein, human
  • Membrane Proteins
  • Mitochondrial Proteins
  • Transforming Growth Factor beta
  • bcl-X Protein