[Effects of the inhibitor of NF-kappaB and IFN-gamma on BAFF-R gene promoter activity].
OBJECTIVE
To investigate the effects of IFN-gamma and the inhibitor of NF-kappaB(BAY11-7082) on BAFF-R gene promoter activity.


METHODS
The fragments with different lengths of the 5'-flanking region of BAFF-R gene were amplified by PCR.Then PCR products were cloned into Luciferase reporter vector (pGL3-Basic) to construct eight recombinant plasmids. These recombinant plasmids were transiently transfected into KM3 cells to locate the promoter region with the highest activity. Then cells transfected with recombinant plasmid with the highest promoter activity were cultured on media in the absence or presence of IFN-gamma and BAY11-7082 and relative luciferase activity were tested and compared. Meanwhile BAFF-R mRNA expression were also examined and compared before and after IFN-gamma and BAY11-7082 were added into cell medium.


RESULTS
IFN-gamma can promote BAFF-R promoter activity and up-regulate BAFF-R mRNA expression. And BAY11-7082 can inhibit BAFF-R promoter activity and down-regulate BAFF-R mRNA expression.


CONCLUSIONS
IFN-gamma and the NF-kappaB pathway could be involved in regulating the transcription and mRNA expression of BAFF-R gene.
Yuan
Hong-xiang
HX
Laboratory Medicine Center; Affiliated Hospital of Nantong University, Medical Key Disciplines of Jiangsu Province, Nantong, China. yhx8284@yahoo.cn
Wang
Yue-guo
YG
Wu
Xin-hua
XH
Ni
Hong-bing
HB
Pu
Jiang
J
Shen
Xian-juan
XJ
Ju
Shao-qing
SQ
D053265:B-Cell Activation Factor Receptor / Q000235:genetics*; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000502:physiology; D009570:Nitriles / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis; D013450:Sulfones / Q000494:pharmacology; D014162:Transfection
C434003:3-(4-methylphenylsulfonyl)-2-propenenitrile; D053265:B-Cell Activation Factor Receptor; D016328:NF-kappa B; D009570:Nitriles; D012333:RNA, Messenger; D013450:Sulfones; C435926:TNFRSF13C protein, human; D007371:Interferon-gamma