Construction of endothelin converting enzyme-1 antisense RNA expression vector from human bronchial epithelial cells
Abstract：Objective To explore the effects of endothelin converting enzyme-1 (ECE-1) antisense RNA on ECE-1 expression in human bronchial epithelial cell line(16HBE). Methods ECE-1 antisense RNA expression vector with green fluorescent protein as a reporter was constructed and transfected into 16HBE with Fugene 6TM.The ECE-1 expression was measured by RT-PCT and Western bloting. Results Agrose gel electrophoresie showed that 0.717kp ECE DNA fragment was inserted into pEGFPC1 plasmid in reverse orientation. After G418 selection, anti-16HBE cell line stably expressing ECE-1 antisense RNA was developed. Both ECE-1 mRNA(ECE/actin β 0.012 5±0.058) and protein expression in anti-16HBE cells were significantly lower than those in normal cells(ECE/actin β 0.134 7±0.067) detected by RT-PCR and Western bloting respectively(P＜0.05). Conclusion ECE-1 expression in 16HBE cells can be inhibited by ECE-1 antisense RNA in vitro.
Machine-generated Keywords：
Publication Date：2001-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 101-103 )