%0 Journal Article
%T Engagement of membrane immunoglobulin enhances Id3 promoter activity in WEHI-231 B lymphoma cells
%A LI Xiao-jun
%A HATA Kikumi
%A MIZUGUCHI Junichiro
%J Acta Pharmacologica Sinica
%D 2016
%B 2016
%9 
%! Engagement of membrane immunoglobulin enhances Id3 promoter activity in WEHI-231 B lymphoma cells
%K 
%X  Aim:  We have recently shown that engagement of membrane immunoglobulin  (mIg) induced upregulation of inhibitor of differentiation 3 (Id3) mRNA, resulting  in growth arrest at G1 phase in WEHI-231 cells. In the present study, we examined  whether engagement of mIg will affect promoter activity of the Id3 gene in WEHI-  231 cells.  Methods:  DNA fragments corresponding to the 5'-flanking region of  mId3 gene were amplified by polymerase chain reaction (PCR) using genomic  DNA as the template. Three DNA fragments upstream of the transcription start  site (+1) of the mId3 gene were subcloned into the luciferase reporter vector PGVB2.  The recombinant constructs were transiently transfected into WEHI-231 cells  by an electroporation method. After incubation for 24 h, WEHI-231 cells were  stimulated with 10 mg/L anti-IgM or irradiated CD40L-expressing NIH3T3 cells or  control NIH3T3 cells for further 24 h, followed by assay for luciferase activity.   Results:  The luciferase analysis demonstrated that basal promoter activity of the  Id3 gene was found in the region between -200 and +54. The Id3 promoter activity  was increased 2-fold following stimulation with anti-IgM, but not CD40L, compared  with medium alone.  Conclusion:  The mIg-mediated upregulation of Id3  expression is controlled, at least in part, through transcriptional regulation, as  assessed by luciferase assay.
%U http://www.chinaphar.com/article/view/3591
%V 26
%N 4
%P 486-491
%@ 1745-7254