白桦脂酸对γδT细胞杀伤胰腺癌细胞功能影响及机制研究  

Research of Mmechanism about Enhancing the Activity of γδT Cell Treated by the Betulinic Acid on Pancreatic Cancer Cell lines SW-1990

作　　者：徐卫东[1] 刘军权[2] 陈复兴[2] 李莹[2] 李凤英[3] 周忠海[2] 费素娟[4] 

机构地区：[1]徐州医学院,221000 [2]中国人民解放军97医院 [3]邹城市中医院 [4]徐州医学院附属医院

出　　处：《长治医学院学报》2013年第3期161-167,共7页Journal of Changzhi Medical College

基　　金：南京军区医学科技创新课题(11MA040)

摘　　要：目的:观察白桦酯酸作用前后对人γδT细胞增殖及对胰腺癌SW-1990细胞杀伤活性的影响,探讨其发生的机制。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γδT细胞,收集培养第7天的γδT细胞,给予不同浓度白桦酯酸诱导48h,CCK8法检测白桦酯酸对γδT细胞增殖的影响,四甲基偶氮唑蓝(MTT)法检测胰腺癌SW-1990细胞增殖率;γδT细胞分泌细胞因子的检测;流式细胞术(FCM)检测白桦酯酸作用前后γδT细胞穿孔素(PFP)、颗粒酶B(GrB)、NKG2D、CD107a的表达变化;乳酸脱氢酶(LDH)释放法测定白桦酯酸对γδT细胞杀伤胰腺癌细胞株SW-1990的活性影响。结果:白桦酯酸浓度在0.013～1.6μg/mL时能促进γδT细胞生长;经白桦酯酸诱导后的γδT细胞PFP、GrB、CD107a、NKG2DIFN-γ和TNF-а的表达显著高于对照组(P<0.05),对胰腺癌SW-1990细胞的杀伤活性亦显著高于对照组(P<0.05)。结论:白桦酯酸在一定浓度范围内能够促进γδT细胞增殖,并增强其对胰腺癌SW-1990细胞的杀伤活性,其机制可能与上调γδT细胞表面PFP、GrB、CD107a、NKG2D的表达和增加γδT细胞IFN-γ和TNF-а分泌有关。Objective ： To observe the effect of betulinic acid （ BetA ） on the growth of humanγδT cells and the killing activity ofγδT cells on the pancreatic cancer cell lines SW-1990in vitro before and after induced by betulinic acid.explore its mechanism.Methods ： Peripheral blood mononuclear cell （ PBMC ） were separated form the healthy and were induced with various of eytokine to becomeγδT cells in vitro.γδT cells were collected on the seventh day and were induced with betulinic acid in different concentrations for 48h , CCK8proliferation assay of γδT cells were treated with BetA.MTT method assay the proliferation rate of cell lines SW-1990.Levels of cytokines produced byγδT cell lines were detected.Flow cytometry （ FCM ） was used to detect the expression changes of perforin , NKG2D , granzyme B and CD107aofγδT cells before and after betulinic acid induced.Lactate dehydrogenase （ LDH ） release assay was used to measure the influence on cytotoxic activity ofγδT cells induced by betulinic acid against pancreatic cancer cell lines SW-1990in vitro.Results ： Betulinic acid can promoteγδT cells growth when the concentration were in 0.013～1.6 μ g / mL , the expression of perforin , NKG2D , granzyme B , CD107a , IFN-γand TNF-аof γδT cells were significantly higher than the control group （ P〈0.05 ） .In the meanwhile , the eytotoxic activity ofγδT cells in vitro against pancreatic cancer cell lines SW-1990were also remarkably higher than the control group （ P〈0.05 ） .Conclusion ： These results suggest that betulinie acid can promoteγδT cells growth in some concentrations and increase the cytotoxic activity ofγδT cells against pancreatic cancer cell lines SW-1990 , its mechanism may related with two factors , on the one hand , increasing expression of CD107a , PFP NKG2D , granzyme B on the surface ofγδT cells , on the ohter hand , enhancing the activetiesγδT cells and accelerating the generation of IFN-γand TNF-а.

关 键 词：白桦酯酸 胰腺癌细胞 ΓΔT细胞 

分 类 号：R392.12[医药卫生—免疫学]