Analysis Of TRAF1 In Hodgkin's Lymphoma Cells

Objective To investigate the function of TRAF1 and CD30-TRAF1 signaling in B cell-derived Hodgkin's lymphoma.Methods Endogenous and CD30 ligand-induced TRAF1 expression at mRNA and protein levels were examined by quantitative RT-PCR and Western blot analyses individualy. RNA interference was preformed to deplete the expression of TRAF1 in L428 cells and to examine the effects of TRAF1 on cellular survival. An ELISA-based NF-κB family transcription factor activity assay was performed to quantify theκB DNA-binding activity in nuclear extracts. The expression of JunB was measuered by Western blot analyses.Results TRAF1 expression was expressed at both mRNA and protein levels in B cell-derived lymphoma cell lines (L428 and KM-H2). CD30 activation via binding to CD30 ligand induced the TRAF1 expression; the relative mRNA expression was increased to 7.26±0.23 from 3.50±0.2, the relative protein expression was increased to 4.53±0.55 from 2.31±0.35. The apoptosis frequency was increased to 27.7±5.8% in TRAF1-deficiency L428 cells compare to 5.7±1.2% in control cells. The p50 and RelA DNA-binding activity were decreased in TRAF1-deficiency L428 cells. The expression of JunB upon CD30 ligand stimulation was not changed in TRAF1-deficiency L428 cells.Conclusions Taken together, the study here gives a better insight into the function of TRAF1 in B cell-derived lymphoma cells. TRAF1 was overexpressed in B cell-derived Hodgkin's lymphoma cells, which was regulated by CD30 signaling pathway. TRAF1 is truly a crucial molecule mediating the activation of the traditional NF-κB activity, which further facilitates the anti-apoptosis.