尼美舒利提高γδT细胞杀伤胃癌细胞的机制探讨  

Mechanistic study of nimesulide on enhancing γδT cell-mediated killing of gastric cancer cells

作　　者：刘军权[1] 陈复兴[1] 巩新建[2] 李慧忠[1] 蔡凯[1] 张宝福[3] 张颂[1] 张娟[1] 

机构地区：[1]解放军第97医院实验科,江苏徐州221004 [2]解放军第97医院普外科,江苏徐州221004 [3]徐州医学院附属医院泌尿外科,江苏徐州221001

出　　处：《细胞与分子免疫学杂志》2010年第3期261-263,266,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：南京军区医学科学技术研究"十一五"计划课题(06MA45)

摘　　要：目的:探讨尼美舒利对人γδT细胞功能影响。方法:按常规方法培养γδT细胞,收集培养第9天的γδT细胞用不同浓度的尼美舒利(分别为0.25、0.5、1、2、4μmol/L)诱导24 h后收集培养上清液检测细胞因子IFN-γ、TNF-α、IL-12,沉淀细胞流式细胞测定穿孔素、粒酶B和NKG2D,LDH法检测γδT细胞杀伤胃癌细胞活性。结果:经尼美舒利诱导后γδT细胞穿孔素、粒酶B表达量(分别为62.8%和72.7%)明显高于对照组(分别为51.4%和60.9%)P<0.05,尤其尼美舒利浓度在1μmol/L时最显著;经尼美舒利诱导后γδT细胞分泌IFN-γ和TNF-α浓度(分别为262.3 ng/L和177.5 ng/L)明显高于对照组(分别为196.1 ng/L和158.5 ng/L)P<0.05;分泌的IL-12浓度与对照组比较无明显差异(P>0.05)。γδT细胞杀伤胃癌细胞SGC-7901和BCG-823的活性在1μmol/L时最高(分别为73%和70%),明显高于对照组(分别为54%和53%)P<0.05。结论:经尼美舒利诱导后γδT细胞的穿孔素、粒酶B含量和γδT细胞杀伤胃癌细胞SGC-7901、BCG-823活性明显高于对照组。其培养上清液中IFN-γ和TNF-α浓度也明显高于对照组。这一结果为临床尼美舒利预防和治疗消化道肿瘤提供了实验依据。AIM：To explore the effect of nimesulide on human γδT cell function.METHODS：γδT cells were cultured routinely,collected on the 9th day,and then induced with nimesulide at indicated concentrations （0.25 μmol/L,0.5 μmol/L,1 μmol/L,2 μmol/L,4 μmol/L,respectively）.Twenty-four hours after induction,the supernatants were collected to detect IFN-γ,TNF-α and IL-12,whereas the cells were assayed for perforin,granzyme B and NKG2D by flow cytometry（FCM） and for killing of gastric cancer cells by LDH.RESULTS：Nimesulide（1 μmol/L） caused γδT cells to express more of perforin and granzyme B（62.8% and 72.7%,respectively）than the control group（51.4% and 60.9%,respectively）（P0.05）.Likewise,nimesulide（1 μmol/L） enabled γδT cells to secret more of IFN-γ and TNF-α（262.3 ng/L and 177.5 ng/L,respectively）than the control group（196.1 ng/L and 158.5 ng/L,respectively）（P0.05）.Nimesulide did not affect IL-12 secreting capability of γδT cells as compared with the control group（P0.05）.Nimesulide-stimulated γδT cells killed more of SGC-7901 and BCG-823 gastric cancer cells（73% and 70%,respectively） than the control group（54% and 53%,respectively）（P0.05）.CONCLUSION：Nimesulide made γδT cells to express more perforin and granzyme B and to secret more IFN-γ and TNF-α into the supernatant,leading to higher killing rate of SGC-7901and BCG-823 gastric cancer cells.The above data provides experimental basis on the clinical use of nimesulide to prevent and treat digestive tract tumors.

关 键 词：γδT细胞 尼美舒利 穿孔素/粒酶B/NKG2D γ-干扰素/α-肿瘤坏死因子/白介素12 胃癌 

分 类 号：R392[医药卫生—免疫学]