关键词: 人β防御素2；基因转染；人膀胱上皮细胞

Abstract:

Objective    To examine expression of humanβ-defensin 2 (hBD2) in human bladder epithelial cells (T24 cells), and to observe the antimicrobial activity of recombinant hBD2 in vitro.  Methods     By using the cationic Liposome-TransFastTM Transfection Reagent, the plasmid vector carrying the hBD2 gene (pCAGGhBD2) was transferred into T24 cells cultured in serum-free medium. Expression of hBD2 mRNA in T24 cells was determined by reverse transcriptionpolymerase chain reaction (RT-PCR). HBD2 peptide expression was detected by Western blot and concentration of recombinant hBD2 in supernatants was measured by ELISA. Antimicrobial activity of recombinant hBD2 was assessed by the colony-forming unit (CFU) assay. Results    hBD2 could be effectively expressed in T24 cells, and the concentration of recombinant hBD2 in supernatants was (36.5±3.2)ng/106 cells. Recombinant hBD2 in supernatants showed antimicrobial activity. Conclusion    The recombinant eukaryotic expressing vector carrying the hBD-2 gene could be effectively expressed in human bladder epithelial cells, and recombinant hBD2 in supernatants has an obvious antimicrobial effect on some bacterial lines.

Key words: Humanβ-defensin 2; Gene transfection; Human bladder epithelial cells