脂多糖对人结肠癌细胞产生β-防御素3的影响

Effect of lipopolysaccharide on expression of beta defensin 3 in human colon cancer RKO cells

目的分析脂多糖(lipopolysaccharide,LPS)对人结肠癌RKO细胞产生的β-防御素3(mouseβ-defensins-3,mBD-3)的影响。方法将不同浓度(0、10、20和40 ng/ml)的LPS作用于RKO细胞0、24、48和72 h,MTT法检测LPS对RKO细胞的最佳抑制浓度;定量RT-PCR及Western blot法检测LPS作用的RKO细胞中mBD-3的表达情况。结果 LPS对RKO细胞的抑制作用在浓度20 ng/ml、培养48 h时最佳。10、20和40 ng/ml LPS作用的RKO细胞,mBD-3 mRNA及蛋白表达水平均明显高于0 ng/ml LPS组,差异有统计学意义(P<0.05)。结论 LPS可以促进mBD-3的表达,为进一步研究mBD-3的产生机制奠定了基础。

Objective To analyze the effect of lipopolysaccharide（LPS）on expression of mouse β-defensin 3（mBD-3）in human colon cancer RKO cells. Methods RKO cells were treated with LPS at various concentrations（0, 10, 20 and 40 ng/ml）for 0, 24, 48 and 72 h separately, based on which the optimal concentration of LPS for inhibition of RKO cells was determined by MTT assay. The expression of mBD-3 in RKO cells was determined by quantitative RT-PCR and Western blot. Results The optimal concentration of LPS for inhibition of RKO cells was 20 ng / ml, while the optimal time for culture was 48 h. Both the expression levels of mBD-3 mRNA and protein in RKO cells treated with 10, 20 and40 ng / ml LPS were significantly higher than those untreated（P〈0. 05）. Conclusion LPS promoted the expression of mBD-3, which laid a foundation of further study on mechanism of mBD-3 production.