Identification of genes responsive to paeoniflorin, a heat shock protein-inducing compound, in human leukemia U937 cells.
OBJECTIVE
Paeoniflorin (PF) isolated from peony root (Paeoniae radix) has been used as a herbal medicine in East Asia for its anti-allergic, anti-inflammatory, and immunoregulatory effects. PF is known to cause apoptosis and to be a chemical heat shock protein (HSP) inducer. With this information, the effects on the gene expression in human leukemia U937 cells treated with PF were investigated.


METHODS
U937 cells, a human myelomonocytic cell line, were treated with PF at different concentrations (0-640 microg/ml). Expression level of Hsp70 was monitored by Western blotting. Gene expression was evaluated using high-density oligonucleotide microarrays and computational gene expression analysis tools and the results were verified by real-time quantitative PCR.


RESULTS
Although cell viability was not affected after PF treatment at a high concentration of 640 microg/ml, PF treatment (80-640 microg/ml) significantly elevated Hsp70 expression in a concentration-dependent manner. When the cells were treated with PF (160 microg/ml; 30 min), 35 up-regulated and 29 down-regulated genes were identified. Among the differentially expressed genes, a significant genetic network containing CDC2, FOSL1 and EGR1 was associated with biological functions such as cell death, gene expression or cellular growth and proliferation.


CONCLUSIONS
The present results indicate that PF affects the expression of many genes including Hsp70 and will provide a better understanding on the molecular mechanism of action of this compound in inducing HSPs in cells.
Salunga
Thucydides L
TL
Department of Diagnostic Pathology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan.
Tabuchi
Yoshiaki
Y
Takasaki
Ichiro
I
Feril
Loreto B
LB
Zhao
Qing-Li
QL
Ohtsuka
Kenzo
K
Tsuneyama
Koichi
K
Kondo
Takashi
T
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001565:Benzoates / Q000494:pharmacology*; D001952:Bridged-Ring Compounds / Q000494:pharmacology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D005960:Glucosides / Q000494:pharmacology*; D018840:HSP70 Heat-Shock Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D039821:Monoterpenes; D020411:Oligonucleotide Array Sequence Analysis; D020298:U937 Cells; D015854:Up-Regulation; D006027:Glycosides
D000894:Anti-Inflammatory Agents, Non-Steroidal; D001565:Benzoates; D001952:Bridged-Ring Compounds; D005960:Glucosides; D018840:HSP70 Heat-Shock Proteins; D039821:Monoterpenes; C015423:peoniflorin; D006027:Glycosides