DC融合疫苗联合CIK细胞对人胃癌细胞凋亡的影响  

Effect of dendritic cell fusion vaccine combined with CIK cells on apoptosis of human gastric cancer cells

作　　者：蒋丽[1] 李辉[1] 张桂英[2] 穆仪冰[1] 刘海涛 Zhang Guiying(Gastroenterology Department of the Fourth Hospital of Changsha,Changsha 410006)

机构地区：[1]湖南省长沙市第四医院消化内科,410006 [2]中南大学湘雅医院消化内科

出　　处：《中国现代医药杂志》2018年第2期1-7,共7页Modern Medicine Journal of China

基　　金：湖南省卫生计生委科研计划课题B类项目(编号:B2016221)

摘　　要：目的研究树突状细胞(DC)融合疫苗与细胞因子诱导杀伤细胞(CIK)共培养对人胃癌细胞凋亡的影响。方法采集胃癌患者外周血单个核细胞(PBMCs)提取诱导生成DC和CIK细胞。聚乙二醇法融合DC与自体胃癌细胞成为融合疫苗。实验分为3组:DC融合疫苗组(简称融合组)、DC组和胃癌组。3组细胞分别与CIK细胞混合培养,流式细胞仪检测CIK细胞亚群CD3^+CD8^+和CD3^+CD56^+细胞表达率。胃癌细胞与3组混合细胞作用后采用Annexin V-FITC/PI染色法观察和检测其凋亡率。Western blot法检测Raf激酶抑制蛋白(RKIP)、NF-κB、Caspase-8蛋白表达。结果胃癌细胞与DC的融合率为26.2﹪,融合DC较未融合DC高表达CD80和CD86(t=74.11、91.84,P<0.05)。与DC组和胃癌组相比,融合组CIK细胞CD3^+CD8^+和CD3^+CD56^+细胞表达率增加(F=60.29、28.68,P<0.05);融合组胃癌细胞凋亡率增加,RKIP、Caspase-8蛋白相对含量表达上调,而NF-κB蛋白相对含量表达下调(F=52.63、21.05、29.89、101.58,P<0.05)。结论DC融合疫苗与CIK细胞共培养后能促进CIK细胞成熟,可能通过上调RKIP、抑制NF-κB从而上调Caspase-8,促进胃癌细胞凋亡,起到抗肿瘤效应。Objective To study the effect of dendritic cell（DC） fusion vaccine on the apoptosis of human gastric cancer cells after co-cultured with cytokine induced killer cells（CIK）. Methods Peripheral blood mononuclear cells（PBMCs） were collected from patients with gastric cancer and induced in DC and CIK cells. Fusion vaccine of DC with autologous gastric cancer cells was generated by polyethylene glycol method. The experiment was divided into three groups ：DC fusion vaccine group, DC group and gastric cancer group. Three groups of cells were separately mixed and co-cultured with CIK cells. Then flow cytometry was used to detect the expression rates of CIK cell subsets CD3~＋CD8~＋ and CD3~＋CD56~＋ cells.The apoptotic rates of gastric cancer cells were observed and analysized by Annexin V-FITC/PI staining methods after reacted with three groups of mixed cells. Western blot method was used to detect the expressions of RKIP,NF-κB and Caspase-8 protein. Results The fusion rate of gastric cancer cells with DC was 26.2﹪. The expressions of CD80 and CD86 in fusion DC were both higher than those in non fusion DC（t=74.11,91.84,P〈0.05）. Compared with DC group and gastric cancer group, the expression rates of CIK cell subsets CD3~＋CD8~＋ and CD3~＋CD56~＋ cells both increased in fusion group（F=60.29,28.68,P〈0.05）. The apoptotic rates of gastric cancer cells were increased, the relative protein expressions of RKIP and Caspase-8 were both up-regulated whereas that of NF-κB was down-regulated in fusion group（F=52.63,21.05,29.89,101.58,P〈0.05）. Conclusion Coculture of DC fusion vaccine and CIK cells can promote the maturation of CIK cells and promote the apoptosis of gastric cancer cell and anti-tumor effect probably by increasing the expression of RKIP,inhibiting the expression of NF-κB and then increasing the expression of Caspase-8.

关 键 词：胃癌 树突状细胞融合疫苗 细胞因子诱导杀伤细胞 凋亡 RAF激酶抑制蛋白 

分 类 号：R735.2[医药卫生—肿瘤]