氢化可的松对人NK细胞增殖及其对胰腺癌SWl990细胞杀伤效率的影响  

Effect of hydrocortisone on proliferation and killing activity of NK cells against SW1990 cells

作　　者：薛成俊[1,2] 周忠海[3] 陈复兴[3] 吕小婷[3] 李莹[3] 费素娟[1] 

机构地区：[1]徐州医学院附属医院消化内科,江苏徐州221002 [2]南通大学附属建湖医院消化内科 [3]解放军第97医院中心实验室

出　　处：《中华胰腺病杂志》2013年第3期179-182,共4页Chinese Journal of Pancreatology

摘　　要：目的探讨氢化可的松对人外周血NK细胞增殖及其对胰腺癌SW1990细胞杀伤力的影响。方法分离健康人外周血单个核细胞加入到含IL-15细胞因子的NK细胞培养基中诱导培养NK细胞。当NK细胞纯度达到70％以上时，加入10^-6、10^-5、10^-4、10^-3μmoL／L氢化可的松继续培养7d。以未加氢化可的松的NK细胞作为对照组。采用锥虫蓝染色计数细胞；采用流式细胞术检测CD3-CD56＋NK细胞含量及其穿孔素、颗粒酶B和IFN-γ的表达；以20：1的效靶比将NK细胞与SWl990细胞共培养，用细胞增殖-毒性检验法（CCK-8）检测NK细胞对SW1990细胞的杀伤效应。结果用氢化可的松处理7d后NK细胞量平均达到70．72％-76．39％，与对照组的（72．61±3．76）％差异无统计学意义；10^-6、10^-5、10^-4、10^-3μmol／L氢化可的松处理组NK细胞的增殖倍数分别为（9．13±0．94）、（9．67±1．51）、（10．33±1．07）、（8．40±1．47）倍，均显著高于对照组的（4．23±0．82）倍（P值均〈0．01）；NK细胞对SWl990细胞的杀伤效率分别为（58．58±4．89）％、（62．27±5．63）％、（64．02±5．79）％、（63．88±3．61）％，均较对照组的（57．46±5．11）％增强，其中10^-4μmol／L组的差异具有统计学意义（P〈0．05）；10^-4、10^-3μmol/L氢化可的松处理组NK细胞穿孔素表达分别为（96．71±3．04）％、（97．56±2．18）％，显著高于对照组的（92．40±3．53）％（P〈0．05或0．01）；10^-5μmol／L氢化可的松处理组NK细胞的颗粒酶B表达为（78．23±2．94）％，显著高于对照组（73．68±3．52）％（P〈0．05）；10^-5、10^-4、10^-3μmol／L氢化可的松处理组NK细胞IFN-γ表达分别为（96．61±2．04）％、（97．58±2．17）％、（98．00±1．77）％，均显著高于对照组的（92．44±2．74）％（P值均〈0．01）。结论氢化可的松可促进IL-15活化的NK细胞增殖，适当浓�Objective To investigate the effects of hydrocortisone （HC） on proliferation and killing activity of NK cells against pancreatic cancer SW1990 cells in vitro. Methods Peripheral blood mononuclear cells of healthy people were isolated and cultured with NK cells medium containing IL-15. When the purity of NK cells reached above 70% , different concentrations of HC （ 10-6, 10-5, 10-4, 10-3μmol/L） were added and co-cultured with NK cells for 7 days. And NK cells without HC were used as control. CD3 - CD56 ± NK cell numbers of each group were countered by trypan blue staining. Perforin, granzyme B and IFN-γ expression of CD3 - CD56＋ NK cells were verified by flow cytometry. NK ceils and SW1990 ceils were co-cultured with a 20：1 effector to target ratio, then the cytotoxie activity of NK cells against SW1990 cells were analyzed by CCK-8 kit.Results After treatment with different concentration of HC for 7 days, NK cells purity of each group reached 70.72% - 76.39%, and it was not significantly different with that in control group [ （72.61 ± 3.76） % ]. The proliferation folds of NK cells treated with 10-6, 10-5, 10-4, 10 3 μmol/L HC were （9.13± 0.94）, （9.67 ± 1.51 ） , （ 10.33 ± 1.07）, （8.40 ± 1.47 ） times, respectively, while it was （4.23 ± 0.82） times in control group （ all P 〈0.01 ）. The killing effects of NK cells on SW1990 cells were （ 58.58 ± 4.89 ） % , （62.27 ± 5.63） % , （64.02 ± 5.79 ） %, （ 63.88 ± 3.61 ） % , which were higher than that in control group [ （ 57.46 ± 5.11 ） %], moreover, the difference between NK cells of 10 -4 μmol/L HC treatment group and control group was statistically significant（P 〈0.05）. The expressions of perforins of 10 4, 10 3 μmol/L HC treatment group were （ 96.71 ± 3.04 ） %, （ 97.56 ± 2.18 ） %, which were significantly higher than that in control group [ （92.40 ± 3.53） %, P 〈 0.05 or 0.01 ]. The expression of granzyme B in 10-5 μmol/L HC treatment group was （78.23 ±2.94）%, which wer

关 键 词：氢化可的松 杀伤细胞 天然 胰腺肿瘤 细胞增殖 杀伤活性 

分 类 号：R735[医药卫生—肿瘤]