[Preliminary study of the molecular regulation of BCG-mediated enhancement of hBD-1 gene expression in human pulmonary gland epithelial cells].
OBJECTIVE
To define the regulation mechanism of the enhancement expression of human beta-defensin-1 (hBD-1) stimulated by bacille Calmette-Guerin (BCG) at transcription level.


METHODS
A series of 5'-flanking deletions of hBD-1 gene were ligated into pEGFP-1 and pCAT basic vector. pEGFP hBD-1 recombinants were transfected into SPC-A-1 cells and the green fluorescence protein (GFP) expression was observed by fluorescence microscopy. SPC-A-1 cells were co-transfected with pCAT hBD-1 constructs and pSV-beta-Galactosidase control vector and were stimulated with effective BCG cell wall components. CAT and beta-Gal protein expressions were determined by ELISA.


RESULTS
The promoting activity of the -69 region was lower than that of the region -575 and -314. After being stimulated with the active fraction of BCG cell wall proteins, the relative CAT expression of pCAT hBD-1/-69 still showed the increasing tendency of -2161 construct. Computer consensus match analysis indicated that the nucleotides between -63 bp to -50 bp was the potential binding site for C/EBP beta.


CONCLUSIONS
The nucleotides from -69 to  bp is essential to the enhancement of hBD-1 gene transcription by BCG.
Zhu
Bingdong
B
Research Unit of Infection & Immunity, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Huang
Ning
N
Wu
Qi
Q
Wang
Boyao
B
D001500:BCG Vaccine / Q000494:pharmacology*; D001483:Base Sequence; D027201:Cationic Amino Acid Transporter 1; D004847:Epithelial Cells / Q000378:metabolism*; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D008168:Lung / Q000166:cytology / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009163:Mycobacterium bovis / Q000737:chemistry; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014158:Transcription, Genetic; D023083:beta-Defensins / Q000096:biosynthesis / Q000235:genetics*
D001500:BCG Vaccine; D027201:Cationic Amino Acid Transporter 1; C406407:DEFB1 protein, human; D012333:RNA, Messenger; D023083:beta-Defensins