胃肿瘤抗原致敏的脐血 DC 联合 CIK 对胃癌细胞株 SGC-7901杀伤活性的研究

Effect of umbilical cord blood dendritic cells induced by gastric cancer antigen combined with CIK cells in gastric cancer cell lines SGC-7901

目的：探讨体外胃肿瘤抗原致敏脐血 DC 联合细胞因子诱导的杀伤细胞（CIK）对胃癌细胞株 SGC-7901的杀伤作用。方法分离脐血单个核细胞培养 DC 细胞和 CIK 细胞。流式细胞仪检测成熟 DC 细胞表面抗原 CD83、CD86、CD11c 及 CIK 细胞表面抗原 CD3、CD56、CD4、CD8、CD16表达。致敏 DC-CIK、非致敏 DC-CIK、CIK 作为效应细胞，SGC-7901作为靶细胞，利用乳酸脱氢酶（LDH）释放法检测致敏 DC-CIK、脐血 DC-CIK、CIK 分别在效靶比10：1、20：1、40：1时对胃癌细胞的杀伤活性。结果成熟脐血 DC 表面抗原 CD83^＋ CD86^＋、CD11c^ ＋ CD83^＋、CD86^＋ CD11c^ ＋表达率分别为（75.4±2.1）％、（79.3±1.4）％、（80.2±2.6）％。致敏脐血 DC 表面表达率分别为（77.7±1.5）％、（82.6±1.9）％、（76.9±2.6）％，二者差异无统计学意义（t ＝1.526，P ﹥0.05；t ＝0.958，P ﹥0.05；t ＝1.049，P ﹥0.05）。成熟 CIK 中 CD4^＋细胞占（22.8±1.3）％，CD8＋细胞占（77.3±1.8）％，CD3^＋ CD56^＋ CD16^＋细胞占（24.5±2.1）％。致敏 DC-CIK、DC-CIK、CIK 都对胃癌细胞有杀伤作用，致敏 DC-CIK 在效靶比为10：1、20：1、40：1时杀瘤活性分别为（37.68±1.49）％、（41.67±0.90）％、（42.71±0.98）％，在效靶比为40：1时杀瘤活性最强，DC-CIK 组分别为（36.77±0.46）％、（38.94±0.95）％、（41.15±0.89）％，CIK组分别为（34.74±1.01）％、（37.76±0.43）％、（39.65±0.79）％，三组间差异有统计学意义（F ＝5.92， P ﹤0.05；F ＝19.13，P ﹤0.05；F ＝8.88，P ﹤0.05）。结论胃肿瘤抗原致敏脐血 DC 可明显增强 DC-CIK的杀瘤活性，致敏脐血 DC-CIK 在效靶比为40：1时杀瘤活性最强。

Objective To investigate the effect of umbilical cord blood dendritic cells（DCs）induced by gastric cancer antigen combined with cytokine induced killer（CIK）cells in gastric cancer cell lines SGC-7901 in vitro. Methods Mononuclear cells from umbilical cord blood were used to create DCs and CIKs. The cell surface antigen expression of the mature DCs such as CD83,CD86,CD11c and the cell surface antigen of CIKs such as CD3,CD56,CD4,CD8,CD16 were detected using flow cytometry. Sensitized DCs-CIKs,DCs-CIKs,CIKs as effective cells,and SGC-7901 as target cells,the killing activities of these effective cells were tested with LDH release,which the number ratio of cells between effective cells and SGC-7901 cells were 10 ：1,20 ： 1,40 ： 1,respectively. Results The cell surface antigen expressions of the mature DCs,such as CD83^ ＋ CD86^ ＋ ,CD11c^ ＋ CD83^ ＋ ,CD86^ ＋ CD11c^ ＋ were（75. 4 ± 2. 1）% ,（79. 3 ± 1. 4）% ,（80. 2 ± 2. 6）% , respectively. The mature sensitive-DCs surface antigen expressions,such as CD83 ^＋ CD86^ ＋ ,CD11c ^＋ CD83^ ＋ , CD86^ ＋ CD11c ^＋ ,were（77. 7 ± 1. 5）% ,（82. 6 ± 1. 9）% ,（76. 9 ± 2. 6）% ,respectively. There was no sta-tistical significance about the surface antigen expression between DCs and sensitive-DCs（t = 1. 526,P ﹥ 0. 05;t = 0. 958,P ﹥ 0. 05;t = 1. 049,P ﹥ 0. 05）. The CIKs surface antigen expressions,such as CD4^ ＋ ,CD8^ ＋ , CD3^ ＋ CD56^ ＋ CD16 ^＋ ,were（22. 8 ± 1. 3）% ,（77. 3 ± 1. 8）% ,（24. 5 ± 2. 1）% ,respectively. The results suggested that the killing effect of the three kinds of combination cells on gastric cancer cells was different. The＆amp;nbsp;number ratio of cells between sensitive-DCs and SGC-7901 cells were 10 ： 1,20 ： 1,40 ： 1,which the killing activities of sensitive-DCs-CIKs against SGC-7901 were（37. 68 ± 1. 49）% ,（41. 67 ± 0. 90）% ,（42. 71 ± 0. 98）% ,respectively. The killing activity of sensitive-DCs-CIKs was the highest when the ratio of cells between sensitive-DCs and SGC-7901 cells were 40 ： 1. The killing activities of DC-CIKs were（36. 77 ± 0. 46）% ,（38. 94 ± 0. 95）% ,（41. 15 ± 0. 89）% ,respectively. The killing activities of CIKs were（34. 74 ± 1. 01）% ,（37. 76 ± 0. 43）% ,（39. 65 ± 0. 79）% ,respectively. There were statistically significant differences among the three groups（F = 5. 92,P ﹤ 0. 05;F = 19. 13,P ﹤ 0. 05;F = 8. 88,P ﹤ 0. 05）. Conclusion The tumor killing activity of CIK is enhanced obviously by umbilical cord blood DCs which is sensitized by gastric cancer tumor antigen. There is the highest killing activity when the number ratio of cells between sensitive-DC-CIK and SGC-7901 cells is 40 ： 1.