Toll like receptor 4 expression and cell immunoregulation in human ovarian cancer cell SKOV3 induced by LPS stimulation / 中华微生物学和免疫学杂志

ABSTRACT

Objective To study the effects of lipopolysaccharide(LPS) on the expression of toll like receptor 4 (TLR4), reactive oxygen species(ROS) and on the proliferation of cells as well as secretion of six proinflammmatory cytokines including TNF-α, IL-1, IL-6, IL-8, IL-10, IL-12 levels in SKOV3 cells. And to explore the mechanism of SKOV3 cells in regulation. Methods Cultured primary SKOV3 cells were stimulated with different concentrations of LPS (0.01 μg/ml, 0.1 μg/ml, 1 μg/ml, 10 μg/ml and 20 μg/ml) for 4 h, the TLR4 expression in SKOV3 cells were examined by flow cytometry;1 μg/ml LPS stimulated SKOV3 for 4 h, 8 h, 12 h, 24 h respectively, the TLR4 expression and cell cycle in SKOV3, cell proliferation, ROS level as well as cells and TNF-α and IL-1, IL-6, IL-8, IL-10, IL-12 levels in the culture medium were assayed by flow cytometry, MTT, CBA assay respectively. Results LPS with different concentrations of LPS stimulation in-duced an increased TLR4 expression, however, the expression was reduced when LPS concentration up to 10 μg/ml. LPS stimulation for 4 h, 8 h induced an increased TLR4 expression and cell proliferation. Stimulated for 24 h, however, the TLR4 expression and cell growth were inhibited in S period. Meanwhile, LPS stimulation for 4 h, 8 h, 12 h, 24 h induced a higher ROS secretion in comparison with control group. LPS stimulation induced a stronger cytokine response in comparison with control group, as demonstrated by the production of TNF-α, IL-1, IL-6, IL-8 secretion in cultured SKOV3 cells, while IL-10 and IL-12 with low expression have no obvious difference in the all medium samples. Conclusion TLR4 expression, cell proliferation, ROS and proin-flammmatory cytokine secretion could be induced in SKOV3 through LPS stimulation. The study provide new ex-periment evidences for human ovarian cells SKOV3 immunity regulation and inflammation reaction to promote cells inhibition after LPS stimulation.