白细胞介素17诱导子宫内膜癌HEC-1-B细胞对单核细胞趋化的研究

The effect of IL-17 treating on HEC-1-B cells to THP-1 chemotaxis

目的:探讨白细胞介素17(IL-17)诱导人子宫内膜癌HEC-1-B细胞对单核细胞的趋化作用,及其对趋化因子CCL2、CCL5表达的影响。方法:分别采用0ng/ml(对照组)、1ng/ml、10ng/ml、100ng/ml浓度的IL-17诱导人子宫内膜癌HEC-1-B细胞,24h后收集细胞上清液置于transwell小室的下室,上室放入THP-1细胞,计数进入下室的THP-1细胞并计算趋化指数;采用荧光定量PCR方法和ELISA方法检测1ng/ml、10ng/ml、100ng/ml IL-17对HEC-1-B细胞CCL2、CCL5 mRNA基因表达以及蛋白表达水平的影响。结果:单核细胞的趋化能力与IL-17呈浓度依赖性,趋化指数在对照组及1ng/ml、10ng/ml、100ng/ml 3个实验组分别为2.05±0.17、2.93±0.22、3.53±0.28及4.57±0.37,4组的差异有统计学意义(P<0.01)。各实验组细胞CCL2及CCL5 mRNA和蛋白表达水平与对照组比较明显升高(P<0.05),表达水平均随着IL-17浓度增加而增高。结论:IL-17诱导后的子宫内膜癌细胞液可以促进单核细胞的趋化作用,其机制可能与IL-17促进HEC-1-B细胞分泌趋化因子CCL2、CCL5的表达上调有关。

Objectives：To investigate the effect of chemotaxis on HEC-1-B cells to THP-1 cells induced by IL-17 and the effect of expression of CCL2 and CCL5.Methods：Transwell assay was performed to examine THP-1 cells chemotaxis induced by IL-17.HEC-1-B cells were cocultured with 0ng/ml（control）,1ng/ml,10ng/ml and 100ng/ml IL-17 respectively.Supernatant were collected after 24 hours and transferred to the lower chambers.THP-1 cells were placed in the upper chambers.The chemotatic index and the number of migrated THP-1 cells in low chamber were counted after 2hours incubation.The expression level of CCL2 and CCL5 mRNA and protein were detected by real-time quantitative PCR and ELISA.Results：The conditioned media from the HEC-1-B cells induced by IL-17 could remarkably stimulate the chemotaxis of THP-1 monocytes in a concentration-related manner.The chemataxis index of four groups were 2.05±0.17,2.93± 0.22,3.53±0.28 and 4.57±0.37 respectively.The differences among four groups were significantly different（P0.01）.The expression level of CCL2,CCL5 mRNA and protein of HEC-1-B cells were higher compared to the untreated cells in a dosage-dependent manner（P0.05）.Conclusion：HEC-1-B cells induced by IL-17 can promote the chemotactic migration of THP-1 cells.The mechanism may be associated with the upregulation of CCL2 and CCL5 expression.