Panax ginseng increases hypoxia-induced down-regulated cellular response related genes in human neuroblastoma cells, SK-N-MC.
BACKGROUND
Many studies have suggested that hypoxia plays a crucial role in the pathogenesis of various neurological disorders. To determine protective effect of Panax ginseng (PG) on hypoxia (0.1% O(2))-induced cell death in human neuroblastoma cells SK-N-MC, we profiled the gene expression among hypoxia, PG-treated hypoxia and normoxia groups.


METHODS
To determine protective effect on hypoxia-induced cytotoxicity of PG, we performed 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. We compared the gene expression profiles among hypoxia, PG-treated hypoxia (100 mug/ml, 6 hours) and normoxia groups using 8K human cDNA microarray analysis. Additionally, in order to identify differentially expressed genes between hypoxia and PG-treated hypoxia groups, hierarchical clustering of genes was also performed.


RESULTS
MTT assay showed that PG protected hypoxia-induced cell death. In cDNA microarray analysis, hypoxia remarkably down-regulated IGF-II mRNA-binding protein 3 (IMP-3), integrin alpha 2 (ITGA2), syndecan binding protein (SDCBP), insulin-like growth factor binding protein 3 (IGBP3) and M-phase phosphoprotein 10 (MPHOSPH10), belonging to category of cellular physiologic response (global M<-3.5). In cluster analysis, 1428 genes exhibited differential expression levels between hypoxia and PG-treated hypoxia groups. Of them, the expressions of 11 genes were increased more than two-fold by PG treatment compared to those in hypoxia group. Particularly, of 11 genes, the expression levels of cellular physiologic response related genes such as MPHOSPH10, IMP-3 and SDCBP, which markedly down-regulated by hypoxia, are increased more than four-fold by PG treatment, compared to hypoxia group.


CONCLUSIONS
In summary, hypoxia induced down-regulation of cellular physiologic response related genes in human neuroblastoma cells, SK-N-MC, and PG ameliorated the hypoxia-induced down-regulation of such genes. These results indicate possible usage of PG in hypoxia-induced neuronal injury including ischemia, trauma and degenerative diseases.
Park
Haejeong
H
Kohwang Medical Research Institute, College of Medicine, Kyung Hee University, Seoul 130-701, Korea.
Kim
Hakjae
H
Ha
Eunyoung
E
Yoon
Seohyun
S
Kim
Mi-Ja
MJ
Hong
Meesuk
M
Leem
Kang-Hyun
KH
Hong
Seung-Jae
SJ
Yang
Jongsoo
J
Chung
Joo-Ho
JH
D000704:Analysis of Variance; D018797:Cell Cycle Proteins / Q000378:metabolism*; D015687:Cell Hypoxia / Q000187:drug effects / Q000502:physiology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects*; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D016547:Kinesins; D009363:Neoplasm Proteins / Q000378:metabolism*; D009447:Neuroblastoma; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D005894:Panax / Q000737:chemistry*; D010750:Phosphoproteins / Q000378:metabolism*; D028321:Plant Preparations / Q000494:pharmacology*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053676:Syntenins / Q000378:metabolism*; D013778:Tetrazolium Salts; D013844:Thiazoles
D018797:Cell Cycle Proteins; C498780:IGF2BP3 protein, human; D009363:Neoplasm Proteins; D010750:Phosphoproteins; D028321:Plant Preparations; D016601:RNA-Binding Proteins; C109621:SDCBP protein, human; D053676:Syntenins; D013778:Tetrazolium Salts; D013844:Thiazoles; C438388:KIF20B protein, human; D016547:Kinesins; C022616:thiazolyl blue