Differential response to lipopolysaccharide by JEG-3 and BeWo human choriocarcinoma cell lines.
OBJECTIVE
To determine the effect of lipopolysaccharide (LPS) on NF-κB gene expression and proinflammatory cytokine release from trophoblast cell models, JEG-3 and BeWo human choriocarcinoma cells.


METHODS
Serum-starved JEG-3 and BeWo cells were treated with LPS (from Escherichia coli serotype 0111:B4) for 24 or 48h. Cell culture medium was collected and assayed for interleukin (IL)-1β, IL-6, IL-8, and transforming necrosis factor (TNF)-α cytokine release using enzyme-linked immunosorbent assays. RNA was extracted from the cells and real-time PCR was performed to measure NF-κB mRNA expression. All results were analyzed by one-way analysis of variance tests followed by Sidak's post hoc analysis. p<0.05 was considered statistically significant.


RESULTS
LPS triggered an inflammatory response in JEG-3 cells by inducing a 1.5-fold increase in NF-κB mRNA expression and TNF-α release (0μg/mL: 15.13±2.14, 1μg/mL: 14.94±0.75, 10μg/mL: 23.05±4.50, p<0.05) and a 2-fold elevation in IL-6 secretion (0μg/mL: 12.54±5.44, 1μg/mL: 25.54±0.91, 10μg/mL: 24.28±4.43, p<0.05). In contrast, BeWo cells were not as sensitive to LPS exposure; NF-κB mRNA expression was unchanged between LPS-treated and control cells, whereas a small but significant 1.3-fold increase in TNF-α release was found (TNF-α: 15.45±1.53pg/mL, control: 12.24±1.00pg/mL, p<0.05). The inflammatory pathways in BeWo cells were found to be active given that treatment of these cells with IL-1β and TNF-α induced IL-6 secretion. Interestingly, 1μg/mL LPS appeared to decrease IL-6 and TNF-α release from BeWo cells. IL-1β and IL-8 secretion were not detected from either cell lines.


CONCLUSIONS
LPS activates the NF-κB pathway in JEG-3 but not BeWo human choriocarcinoma cells and this may be the reason for their differential inflammatory response to LPS exposure.
UQ Centre for Clinical Research, The University of Queensland, Herston, Queensland, Australia.
Yong Q
YQ
Koh
UQ Centre for Clinical Research, The University of Queensland, Herston, Queensland, Australia.
Hsiu-Wen
HW
Chan
UQ Centre for Clinical Research, The University of Queensland, Herston, Queensland, Australia.
Marloes Dekker
MD
Nitert
UQ Centre for Clinical Research, The University of Queensland, Herston, Queensland, Australia.
Kanchan
K
Vaswani
UQ Centre for Clinical Research, The University of Queensland, Herston, Queensland, Australia.
Murray D
MD
Mitchell
UQ Centre for Clinical Research, The University of Queensland, Herston, Queensland, Australia. Electronic address: g.rice@uq.edu.au.
Gregory E
GE
Rice
D045744:Cell Line, Tumor; D016207:Cytokines / Q000378:metabolism*; D006801:Humans; D008070:Lipopolysaccharides; D016328:NF-kappa B / Q000378:metabolism*; D014327:Trophoblasts / Q000276:immunology* / Q000378:metabolism
D016207:Cytokines; D008070:Lipopolysaccharides; D016328:NF-kappa B
Cytokine; Inflammation; Lipopolysaccharide; Trophoblast