摘要
目的探讨β-谷甾醇对用多种细胞因子和刺激分子联合诱导培养的人外周血单个核细胞（共刺激细胞）杀伤胃癌SGC-7901细胞的影响及其机制。方法分离健康者外周血单个核细胞（PBMC）在体外经多种细胞因子诱导为共刺激细胞,不同质量浓度的β-谷甾醇分别作用于共刺激细胞及胃癌SGC-7901细胞24、48、72 h后,CCK8法检测共刺激细胞的生长,MTT法检测胃癌SGC-7901细胞的生长,FCM检测共刺激细胞PFP、GraB、CD107a、IFN-γ的表达;LDH释放法检测共刺激细胞对SGC-7901细胞的杀伤活性;Western blot检测共刺激细胞中p-ERK1/2、Bcl-2的表达情况。结果培养10 d后,共刺激细胞增殖倍数达高峰。与对照组比较,质量浓度2.5~10μg/ml的β-谷甾醇作用后共刺激细胞的增殖率显著提高（P〈0.05）,质量浓度10~80μg/ml的β-谷甾醇对SGC-7901细胞生长抑制率显著提高（P〈0.05）,并且0.3~5μg/ml的β-谷甾醇作用后的共刺激细胞对SGC-7901细胞的杀伤活性明显增强（P〈0.05）,共刺激细胞中PFP、GraB、IFN-γ及p-ERK1/2、Bcl-2的表达较对照组显著增加（P〈0.05）。结论β-谷甾醇在一定质量浓度范围内能够促进共刺激细胞的增殖,并增强其对胃癌SGC-7901细胞的杀伤活性,其机制可能与β-谷甾醇上调共刺激细胞PFP、GraB、IFN-γ的表达,活化p-ERK1/2、Bcl-2有关。
The study designed to explore the effect of β-sitosterol on killing activity of costimulatory cells（peripheral blood mononuclear cell cocultured with various cytokines and stimulating molecules） against gastric cancer SGC-7901 cells. Peripheral blood mononuclear cells（PBMC） were separated form healthy donors, and then induced with various cytokines to prepare costimulatory cells in vitro. After cultivation with different concentrations of β-sitosterol for 24, 48, 72 h, costimulatory cells and gastric cancer SGC-7901 cells were detected by CCK8assay and MTT assay separately for testing the growth of these two cells. Simultaneously, FCM was used to test the expression of PFP, GraB, CD107a and IFN-γ of costimulatory cells; LDH method was used to test the cytotoxic activity of costimulatory cells against gastric cancer cells; Western blot was used to test the p-ERK1/2 and Bcl-2expression of costimulatory cells. Data indicated that the proliferation multiple of costimulatory cells reached a peak at the tenth day after cell culture. Compared with control group, the proliferation ratio of costimulatory cells increased markedly after cultured with β-sitosterol at concentrations of 2.5-10 μg/ml（P 0.05）. Furthermore, β-sitosterol could inhibit the growth of gastric cancer SGC-7901 cell obviously at concentrations of 10-80 μg/ml（P 0.05）. In addition, β-sitosterol（0.3-5 μg/ml） could significantly enhance the killing activity of costimulatory cells to SGC-7901 cells（P 0.05）, and the expression of PFP, GraB, IFN-γ, p-ERK1/2, andBcl-2 on costimulation cells increased significantly（P 0.05）, as compared with controls. In conclusion, β-sitosterol at appropriate concentration could promote costimulatory cells proliferation and enhance the killing activity of costimulatory cells against gastric cancer cells, and the mechanism may involve in up-regulation of PFP, GraB and IFN-γ expression and activation of the p-ERK1/2 and Bcl-2 signaling pathways.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2014年第7期578-584,共7页
Immunological Journal
