Abstract: Objective：To construct the stable transfection Hela cell lines with lentiviral vector which may interfer the expression of ALDH-1，and to further explore the biological characteristics between the expression of ALDH-1 and Hela cell lines. Methods：Hela cells were cultured in vitro，added the different concentration of puromycin to screen，，and to determine the best screening concentratioｎ；Hela cells were tested from 24-well plates，threw away the old culture medium， and added 400 μL of new medium to the plates，and then added four kinds of lentivirus solution（ALDH-1， ALDH1-1719， ALDH1-740， ALDH1-921，and there was a kind of lentivirus which has the best interfering effect） and one kind of negative control lentivirus solution （LV3-NC） to the plates， at last added Polybrene liquid to them. Every lentivirus solution volume was added about 50 μL （each of viral titers were 1×108 TU/mL），and the Polybrene was added about 0.5 μL （concentration of 5 mg/L） respectively. Fresh culture medium was changed after 24 hours，and observe the fluorescence after 72 hours. At last，using the best screening concentration of puromycin to screen the transfection Hela cells，and to get the stable transfection cell lines. Results：The best screening concentration of puromycin was 0.9 mg/L，lentiviral vectors were successfully transfected into Hela cells，and the stable transfected cell lines were successfully obtained after two weeks. Conclusions：Hela cells which has stable interference of ALDH-1 were successfully constructed.

Key words: Oxidoreductases, Acetaldehyde, Lentivirus, Genetic vectors, Transfection, Hela cells