荷载CSCs抗原的DC疫苗联合小剂量TP治疗乳腺癌小鼠效果观察  

Efficacy of CSCs antigens-loaded dendritic cell vaccine combined with low-dose TP in treatment of breast cancer in mice

作　　者：陈心[1] 梁慧玲[1] 伍龙[1] 徐细明[1] 张一桥[1] 陈能[1] 冯翎[1] 

机构地区：[1]武汉大学人民医院,武汉430060

出　　处：《山东医药》2017年第41期16-19,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81302133);中央高校基本科研业务费专项基金资助项目(2042014kf0149);湖北省自然科学基金资助项目(2012FKC143)

摘　　要：目的研究荷载肿瘤干细胞(CSCs)抗原的树突状细胞疫苗联合小剂量TP(紫杉醇+顺铂)治疗小鼠乳腺癌的疗效。方法利用ALDEFLUOR染料从小鼠乳腺癌细胞系4T1中分选出ALDH high肿瘤干细胞,将CSCs制备成细胞裂解液加载在DC细胞上制备成CSCs-DC疫苗。利用小鼠乳腺癌细胞系4T1接种Balb/c小鼠建立小鼠乳腺癌模型,随机分成磷酸盐缓冲液组(PBS组)、紫杉醇+顺铂组(TP组)、CSCs-DC疫苗组(CSCs-DC组)、CSCs-DC疫苗联合TP组(CSCs-DC+TP组),PBS组腹腔注射0.1 m L的PBS;TP组腹腔注射紫杉醇20 mg/kg,顺铂5 mg/kg;CSCs-DC组皮下注射CSCs-DC疫苗;CSCs-DC+TP组皮下注射CSCs-DC疫苗,腹腔注射紫杉醇20 mg/kg,顺铂5 mg/kg。观测各组肿瘤大小和小鼠生存期,实验终点取小鼠脾脏,体外培养激活后利用LDH细胞毒性实验测定细胞毒性T淋巴细胞(CTL)活性,ELISA法测定B细胞分泌Ig G水平。结果 CSCs-DC+TP组肿瘤体积小于PBS组、TP组及CSCs-DC组(P均<0.05)。小鼠生存期CSCs-DC+TP组最长,生存时间为(62.00±2.65)d,CSCsDC组次之,生存时间为(52.00±1.00)d,TP组和PBS组生存时间分别为(46.00±2.00)、(35.00±1.00)d。CSCsDC+TP组小鼠脾脏中CTL活性和B细胞分泌Ig G水平最高,CSCs-DC组次之,TP组较CSCs-DC组和CSCs-DC+TP组低,但较PBS组高,差异有统计学意义(P均<0.01)。结论 CSCs-DC疫苗联合小剂量TP治疗乳腺癌小鼠较CSCs-DC疫苗和TP单用效果好。Objective To study the therapeutic effect and mechanism of cancer stem cells（ CSCs） antigens-loaded dendritic cell（ DC） vaccine combined with low-does TP（ paclitaxel and cisplatin） in treatment of breast cancer in mice.Methods We sorted out the CSCs from breast cancer cell lines（ 4 T1） by using ALDEFLUOR. Bone marrow-derived DCs were pulsed with CSCs lysate to generate CSCs-DC vaccine. The Balb/c mice were inoculated with the heterogeneous 4 T1 tumor cells by subcutaneous injection to make the mouse breast cancer models. The models were then divided into four groups： PBS group which was administrated PBS,TP group which was injected with 20 mg/kg paclitaxel and 5 mg/kg cisplatin intraperitoneally,CSCs-DC vaccine group（ CSCs-DC group） which was administrated CSCs-DC vaccine by subcutaneous injection,and the CSCs-DC vaccine ＋ TP group（ CSCs-DC ＋ TP group） which was administrated CSCs-DC vaccine by subcutaneous injection and was injected with 20 mg/kg paclitaxel and 5 mg/kg cisplatin intraperitoneally. Tumor volumes were measured and survival was monitored. At the end of the experiment,the spleens in mice were harvested,the cytotoxic lymphocyte（ CTL） activity was measured by using LDH cytotoxicity assay,and Ig G levels secreted by B cells was measured by ELISA. Results The tumor size in the CSCs-DC ＋ TP group was less than that of the PBS group,the TP group and the CSCs-DC group（ P〈0. 05）. The average survival time of the CSCs-DC ＋ TP group was（ 62. 00 ± 2. 65） d and was the longest,the average survival time of CSCs-DC group was（ 52. 00 ± 1. 00） d,the average survival time of TP group and PBS group were（ 46. 00 ± 2. 00） d and（ 35. 00 ± 1. 00） d,respectively. The CTL activity and Ig G levels secreted by B cells in the CSCs-DC ＋ TP group were the highest,the CSCs-DC group was the second,TP group was lower than the CSCs-DC group and the CSCs-DC ＋ TP group,but was higher than the PBS group,and the difference was statistically significant（ P 〈0. 01）

关 键 词：乳腺肿瘤 肿瘤干细胞 疫苗 紫杉醇 顺铂 小鼠 

分 类 号：R737.9[医药卫生—肿瘤]