K-ras突变多肽负载的DC细胞增强CIK细胞对胰腺癌细胞的杀伤作用  

Killing effect of CIKs on pancreatic cancer cells enhanced by DCs loaded with K-ras mutant peptide

作　　者：窦春鹏[1] 李奎武[1] 谭广[2] 

机构地区：[1]辽宁省阜新市中心医院普外一科,辽宁阜新123000 [2]大连医科大学附属第一医院普外二科,辽宁大连116011

出　　处：《中国普通外科杂志》2015年第3期357-362,共6页China Journal of General Surgery

基　　金：国家自然科学基金资助项目(81071173)

摘　　要：目的:观察经K-ras(12-Val)突变多肽负载的树突状细胞(DC)与细胞因子诱导的杀伤细胞(CIK)共培养以后对胰腺癌PANC-1细胞的杀伤作用。方法:取健康人外周血体外诱导分别扩增出DC和CIK;用K-ras突变体抗原表位肽负载DC(K-rasDC),将单纯DC或K-ras-DC与CIK共培养,获得DC-CIK或K-ras-DC-CIK。比较CIK与K-ras-DC-CIK的增殖活性;分别分析DC与K-ras-DC以及CIK与K-ras-DC-CIK的免疫表型差异;检测CIK、DCCIK、K-ras-DC-CIK上清液中IFN-γ、IL-12的水平;检测K-ras-DC-CIK、DC-CIK、CIK对PANC-1细胞的体外杀伤力。结果:K-ras-DC-CIK的增殖能力明显强于单纯CIK(P<0.05);K-ras-DC的成熟表面分子CD1a、CD80、CD83、HLA-DR的表达明显高于单纯DC,而K-ras-DC-CIK细胞群的CD3+CD8+、CD3+CD56+表达率明显高于单纯CIK细胞群(均P<0.05);上清液中IFN-γ、IL-12的水平以及对PANC-1细胞的杀伤力由高到低均依次为K-ras-DC-CIK、DC-CIK、单纯CIK(均P<0.05)。结论:K-ras突变多肽负载后能促进DC的成熟,负载K-ras突变多肽后的DC能增加CIK的增殖及对胰腺癌细胞的杀伤作用。Objective：To observe the killing effect of the cytokine induced killer cells（CIKs） after co-culture with dendritic cells（DCs） harboring K-ras（12-Val） mutant peptide on pancreatic cancer PANC-1 cells.Methods：DCs and CIKs were induced and enriched from peripheral blood of healthy donors,respectively.DCs were loaded with the K-ras mutant epitope peptide（K-ras-DCs）,and CIKs were co-cultured with un-loaded DCs or K-ras-DCs to obtain the DC-CIKs and K-ras-DC-CIKs,respectively.The proliferative activities between CIKs and K-ras-DC-CIKs were compared,the difference in immunophenotype between DCs and K-ras-DCs as well as between CIKs and K-ras-DC-CIKs were analyzed,the IFN-y and IL-12 levels in the culture supernatants from CIKs,DC-CIKs and K-ras-DC-CIKs were measured,and the killing abilities of CIKs,DC-CIKs and K-ras-DCCIKs on PANC-1 cells in vitro were determined.Results：The proliferative ability of K-ras-DC-CIKs was significantly greater than that of the untreated CIKs（P〈0. 05）;the expressions of the mature surface proteins that included CD1 a,CD80,CD83 and HLA-DR in K-ras-DCs were significantly higher than those in un-loaded DCs,while the expression rates of CD3~＋CD56~＋ and CD3~＋CD8~＋ in K-ras-DC-CIK cell population were significantly higher than those in pure CIK population（all P〈0. 05）;the levels of IFN-y and IL-12 in the cell culture supernatant,and the killing ability on PANC-1 cells from high to low order were K-ras-DC-CIKs,DC-CIKs,and pure CIKs（all P〈0. 05）.Conclusion：K-ras mutant peptide can promote DCs maturation,and DCs harboring K-ras mutant peptide can increase the proliferation of CIKs and killing effect on pancreatic cancer cells.

关 键 词：胰腺肿瘤 树突细胞 杀伤细胞 淋巴因子激活 

分 类 号：R735.9[医药卫生—肿瘤]