摘要
目的探讨环磷酰胺对人γδT细胞杀伤胃癌细胞SGC-7901作用的影响。方法异戊烯焦磷酸法体外扩增人外周血γδT细胞,不同浓度的环磷酰胺诱导γδT细胞24 h,四甲基偶氮唑蓝（MTT）法检测环磷酰胺对γδT细胞增殖率的影响,LDH法检测γδT细胞对胃癌细胞的杀伤活性,流式细胞仪检测诱导前后的γδT细胞的凋亡率及NKG2D受体表达水平。结果γδT细胞培养10天时从扩增前4.21%增加到70.35%,当环磷酰胺浓度在0.05~2.0 ng/L时能够促进人γδT细胞的生长,增殖率随着该药物浓度的增高而增高,浓度大于2.0 ng/L时抑制γδT细胞的生长,γδT细胞杀伤活性与对照组比较明显增高,凋亡率与对照组比较明显降低,且γδT细胞表面NKG2D表达水平明显高于对照组。结论环磷酰胺在一定的浓度范围内,促进γδT细胞的增殖,提高γδT细胞对胃癌细胞的杀伤活性,抑制γδT细胞的凋亡,增强γδT细胞表面NKG2D的表达水平,为环磷酰胺在肿瘤免疫治疗中提供了一定的理论依据。
Objective To explore the effect of cyclophosphamide （CTX） on the role of human γδT cells in killing gastric cancer cell line SGC-7901.Methods γδT cells from human peripheral blood was amplified in vitro using isopentenyl pyrophosphate,and was induced at various concentrations of CTX for 24 hours.MTT assay was used to detect the effect of CTX on the proliferation rate of γδT cells.LDH assay was used to detect the cytotoxic activity of γδT cells.Flow cytometry was performed to determine the apoptosis rate of γδT cells and the expression of NKG2D receptor before and after CTX induction.Results The proliferation rate of γδT cells increased from 4.21% to 70.35% at the end of ten days of cultivation.CTX at concentrations of 0.05-2.0 ng/L could promote the growth of γδT cells,with a positive correlation between cell proliferation and drug concentrations,while CTX at concentrations over 2.0 ng/L inhibited the cell growth.There was a marked increase in cytotoxic activity and an evident decrease in cell apoptosis as well as an upregulated expression level of NKG2D on the surface of γδT cells,as compare to the control group.Conclusion CTX at concentrations within a certain range has effect on γδT cells,such as promoting the cell proliferation,enhancing the cytotoxic activity on gastric cancer cells,inhibiting the cell apoptosis and enhancing the expression of NKG2D on the surface of γδT cells.These results may provide theoretical basis for the application of CTX in tumor adoptive immunotherapy.
出处
《徐州医学院学报》
CAS
2010年第6期402-405,共4页
Acta Academiae Medicinae Xuzhou
关键词
环磷酰胺
ΓΔT细胞
胃癌细胞
cyclophosphamide
γδTcells
gastric cancer cells
