[LRP16 gene function based on bioinformatic analysis].
OBJECTIVE
LRP16 is a human novel gene linked to leukemia identified recently. However, its biological function is not fully clarified so far. This study was to investigate the biological function of human LRP16 gene by database-aided bioinformatics analysis.


METHODS
The structures and functions of LRP16 gene promoter and its coding protein were analyzed using bioinformatics prediction, and further experimental testing was performed. The recombinants of pGL3-basic and LRP16 promoter subclones were constructed for luciferase activity analysis. The recombinant of LRP16 open reading frame coding sequence and pcDNA3.1 eukaryotic expression vector was established and transfected into HL-60 and K562 cell lines. DNA damage of HL-60 cells after ultraviolet irradiation was evaluated using single cell gel electrophoresis. Cell cycle of K562 cells was analyzed by flow cytometry.


RESULTS
LRP16 promoter was a typical class II eukaryotic promoter and its core regulation sequence was located within upstream -600 bp of transcriptional start site. In addition, seven cis-acting elements, which may be implicated in cell cycle, hematopoiesis regulation, cell proliferation and repair of DNA damage, were identified. Long type LRP16 coding protein contained homologous sequences of hismacro, COG2110, and A1pp with human histone H2A1C between 148 and 315 amino acid residue. The number of comet cells and the length of comet tail in HL-60 cells irradiated were significantly decreased and the number of living cell was significantly increased in LRP16-overexpression group compared with empty plasmid control group. The proliferation rate and ratio or quantity of G2/M and S phases were significantly increased in LRP16-overexpression K562 group compared with empty plasmid control group. LRP16-overexpression in K562 cells promoted the transition of G1 to S phase and plateau phase of cell proliferation was advanced.


CONCLUSIONS
Promoter regulation prediction and protein domain analysis based on bioinformatics contribute to the study of gene function. LRP16 may play an important role in leukemia progression by promoting cell proliferation, regulating cell cycle, and antagonizing radiation-induced DNA damage.
Yang
Bo
B
Department of Geriatric Hematology, PLA General Hospital, Beijing 100853, P. R. China.
Lu
Xue-Chun
XC
Chi
Xiao-Hua
XH
Han
Wei-Dong
WD
Yu
Li
L
Lou
Fang-Ding
FD
D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology*; D002265:Carboxylic Ester Hydrolases; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D019295:Computational Biology; D004249:DNA Damage; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D018922:HL-60 Cells; D006801:Humans; D020014:K562 Cells; D008297:Male; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D011401:Promoter Regions, Genetic; D014162:Transfection
D009363:Neoplasm Proteins; D002265:Carboxylic Ester Hydrolases; C476832:MACROD1 protein, human