[Establishment and characterization of the lung cancer A549 cell stains with down-regulated expressed HOGG1 genes].
OBJECTIVE
To establish lung cancer A549 cell strains with down-regulated expressed HOGG1 genes and study its biological characteristics.


METHODS
The eukaryotic expression vectors with genes of hammerhead ribozyme targeting human 8-oxoguanine DNA glycosylase 1 (HOGG1) mRNA were transfected into A549 cells by liposmes, and then were screened by G418 till stable cell strains were constructed. The positive recombinants were identified by RT-PCR to amplify NEO genes. The efficacy of inhibition was tested by RT-PCR. Furthermore, morphology, survival curves, cell cycle, cloning efficiency grow in the soft agar and the activity of superoxide dismutase (SOD) were investigated.


RESULTS
The HOGG1-down-regulated expressed A549 cell strains were obtained after G418 selection and NEO genes identification. The mRNA expression level of HOGG1 genes in the down-regulated expressed cells named A549-R were decreased 61.5% than in the untransfected cells A549 (P < 0.05). There was no significant difference between A549-R cells and A549 cells in morphology, the population doubling time and cell cycle. The numbers of clones in the soft agar of A549-R cells was decreased nearly to 50% than of A549 cells (P < 0.05), but the activity of SOD was increased obviously (P < 0.05).


CONCLUSIONS
The lung cancer A549 cell strains with down-regulated expressed HOGG1 genes by silencing and incision of ribozyme were successfully established. Its biological characteristics have no significant changes in most of indexes.
Department of Environmental Health, School of Public Health, Sichuan University, Chengdu 610041, China.
Zun-zhen
ZZ
Zhang
Qin
Q
Zhang
Na
N
Li
Zheng-chang
ZC
Heng
D000230:Adenocarcinoma / Q000235:genetics / Q000473:pathology; D045744:Cell Line, Tumor; D045647:DNA Glycosylases / Q000235:genetics* / Q000378:metabolism; D004260:DNA Repair / Q000235:genetics; D015536:Down-Regulation; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000473:pathology; D018384:Oxidative Stress; D016337:RNA, Catalytic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D016337:RNA, Catalytic; D012333:RNA, Messenger; C427204:hammerhead ribozyme; D045647:DNA Glycosylases; C479058:oxoguanine glycosylase 1, human