热休克胃癌细胞负载DC-CIK细胞杀伤癌细胞活性的研究

检验医学 ›› 2012, Vol. 27 ›› Issue (10): 849-853.

热休克胃癌细胞负载DC-CIK细胞杀伤癌细胞活性的研究

张成磊,陈耀平,杨宝珍,吴若芬,王妍柏

1.宁夏医科大学，宁夏银川 750004；2.宁夏医科大学总医院医学实验中心，宁夏银川 750004

收稿日期:2012-05-14 修回日期:2012-07-05 出版日期:2012-10-30 发布日期:2012-10-12
通讯作者: 杨宝珍，联系电话：0951-6746955。
作者简介:张成磊，男，1984年生，学士，主要从事临床检验诊断学研究。
基金资助:
宁夏自然科学基金资助项目（NZ09111）

Study on the killing activity of DC-CIK cells pulsed with gastric cancer cells of heat shock against cancer cells

1. Ningxia Medical University，Ningxia Yinchuan 750004，China；2. Medical Research Center，Ningxia Medical University General Hospital，Ningxia Yinchuan 750004，China

Received:2012-05-14 Revised:2012-07-05 Online:2012-10-30 Published:2012-10-12

摘要/Abstract

摘要： 目的　研究负载相关抗原的树突状细胞（DC）联合细胞因子诱导的杀伤（CIK）细胞对胃癌细胞的杀伤作用。方法　取健康人外周血并分离出外周血单个核细胞（PBMC），经不同细胞因子分别诱导出DC和CIK细胞，热休克方法处理胃癌MKN-28细胞株并制备肿瘤抗原用于负载DC，与CIK细胞共培养，以流式细胞术检测DC、CIK细胞表型，MTS法检测CIK细胞对MKN-28细胞的杀伤作用。结果　CIK与DC共培养后，与单纯CIK细胞相比较，增殖活性明显提高，CD3⁺CD8⁺、CD3⁺CD56⁺双阳性细胞数增多并且具有更强的杀瘤活性，效靶比为20.0∶1时，负载MKN-28抗原的DC-CIK（Ag-DC-CIK）组对MKN-28的杀伤活性为（57.96±2.23）%，远大于未负载MKN-28抗原的DC-CIK组[（38.02±2.06）%]和单纯CIK组[（29.78±1.84）%]，差异具有统计学意义（P＜0.05）。结论　以热休克凋亡肿瘤细胞抗原负载的DC能促进CIK细胞增殖分化，并提高其对胃癌细胞的杀伤活性。

关键词: 细胞因子诱导的杀伤细胞, 树突状细胞, 热休克, 胃癌

Abstract: Objective　To study the killing effect of dendritic cells (DC) and cytokine-induced killer (CIK) cells pulsed with specific antigens against gastric cancer cells. Methods　Peripheral blood mononuclear cells（PBMC）were isolated from healthy subjects. DC and CIK cells were induced by different cytokines. Gastric cell line MKN-28 lysates of heat shock were used as antigen and pulsed DC，which co-cultured with CIK cells. The phenotypes of DC and CIK cell membranes were determined by flow cytometry，and the killing activity of CIK against MKN-28 was determined by (MTS). Results　Compared with CIK，the co-cultured DC-CIK presented a significantly higher proliferation. The numbers of CD3⁺CD8⁺ and CD3⁺CD56⁺ cells increased，and the killing effect was enhanced. The cytotoxic activity to MKN-28 of DC-CIK pulsed with MKN-28 antigen(Ag-DC-CIK) group was （57.96±2.23）% ，which was more strong than（38.02±2.06）% of DC-CIK pulsed without MKN-28 antigen group and（29.78±1.84）% of CIK group，respectively [the ratio of effect to target =20.0∶1]. Conclusions　Heat shock tumor-lysate-pulsed DC can strengthen the proliferation and killing activity of CIK against gastric cancer cells.

Key words: Cytokine-induced killer cell, Dendritic cell, Heat shock, Gastric cancer

张成磊1，陈耀平2，杨宝珍2，吴若芬2，王妍柏1. 热休克胃癌细胞负载DC-CIK细胞杀伤癌细胞活性的研究[J]. 检验医学, 2012, 27(10): 849-853.

ZHANG Chenglei 1，CHEN Yaoping 2，YANG Baozhen 2，WU Ruofen 2，WANG Yanbai 1. Study on the killing activity of DC-CIK cells pulsed with gastric cancer cells of heat shock against cancer cells[J]. , 2012, 27(10): 849-853.

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