耐药乳腺癌裸鼠模型免疫治疗初探  

Primary Research of Immunotherapy on Drug-Resistance Breast Cancer Bearing Mouse Model

作　　者：庞华[1] 司玉玲[1] 綦振家[1] 王英娟[1] 王亮[1] 李世俊[1] 

机构地区：[1]天津市第四中心医院肿瘤血液科,300140

出　　处：《天津医药》2011年第12期1136-1140,共5页Tianjin Medical Journal

基　　金：天津市自然科学基金支持项目(项目编号:09JCYBJC10600)

摘　　要：目的:观察负载耐药乳腺癌细胞抗原的树突细胞(DC)与细胞因子诱导的杀伤细胞(CIK)联合培养后,用于治疗耐药乳腺癌裸鼠模型的抑瘤效果。方法:分离健康人外周血获得单个核细胞,分别诱导为DC和CIK细胞。将人类乳腺癌耐药细胞株MCF-7/ADR细胞接种裸鼠建立荷瘤裸鼠模型。利用MCF-7/ADR细胞的冻融物抗原冲击DC(AP-DC),然后将AP-DC和抗原未冲击DC分别与CIK细胞共培养(AP-DC+CIK、DC+CIK),并设立CIK细胞单独培养组(CIK)和生理盐水组,将上述4组细胞经尾静脉注入荷瘤裸鼠体内,观察裸鼠肿瘤体积变化,根据瘤质量计算抑瘤率。用流式细胞仪检测DC和CIK细胞成熟表型,酶联免疫吸附法(ELISA)测定各组CIK细胞分泌肿瘤坏死因子(TNF)-α、干扰素(IFN)-γ和白细胞介素(IL)-2的水平。结果:DC和CIK细胞共培养后,2种细胞成熟表型均增高,共培养后的CIK细胞分泌IFN-γ、TNF-α和IL-2的能力亦得到提高,以上均以AP-DC+CIK组增高最为明显;荷瘤裸鼠经上述4组效应细胞治疗后,对照组肿瘤进一步增大,治疗组生长受到抑制,肿瘤体积不同程度缩小,肿瘤质量亦存在差异,以AP-DC+CIK组肿瘤生长明显受抑制,瘤体质量最小,抑瘤率最高。结论:经耐药肿瘤抗原负载的DC与CIK共同作用后,对同种耐药肿瘤的裸鼠抑瘤能力最强。Objective： To investigate the tumor-inhibitory effect of cytokine-induced killer cells （CIK） co-cultured with dendritic cells （DC） loaded multidrug resistance （MDR） breast cancer antigen on the same MDR mammary tumor-bearing mice. Methods： DC and CIK cells were respectively derived from peripheral blood mononuclear cells （PBMC） of healthy individuals. MCF-7/ADR, a kind of breast cancer cell line expressed multidrug resistance, was vaccinated to nude mice to set up tumor bearing mice model. MCF-7/ADR also was prepared to obtain the antigen lyses. CIK was co-cultured with DC pulsed or unpulsed by the above antigen lyses （AP-DC＋CIK and DC＋CIK）. The tumor-bearing mice were injected by intravenous with AP-DC＋CIK, DC＋CIK, CIK and normal saline （NS）. The antitumor effects were evaluated and compared in 4 groups through the tumor size, weight and the tumor inhibition rate. And the cell phenotypes of DC and CIK were analyzed by flow cytometry. The secretion levels of TNF-α, IFN-γ and IL-2 were analyzed by ELISA in 4 groups. Results： The mature phenotypes of two kinds of cells （DC and CIK） were increased, and secretion levels of IFN-γ, TNF-α and IL-2 were increased in CIK cells after co-cultured with DC. There was the highest increase level in AP-DC＋CIK group than that of other 3 groups. It was found that the tumor size was accreted in NS group, and was inhibited in the other 3 tumor-bearing mouse groups after treated with the effector cells. The tumor weights were also different in treatment group and control group. There was the highest inhibition on tumor growth and tumor inhibition rate in AP-DC＋CIK group than that of other groups. Conclusion： There was a higher tumor-inhibitory effect after CIK cells were co-cultured with DC loaded with multidrug resistance tumor antigen.

关 键 词：树突细胞 乳腺肿瘤 癌 P糖蛋白 杀伤细胞 肿瘤 实验性 小鼠 SCID模型 动物 

分 类 号：R28[医药卫生—中药学]