树突细胞/肿瘤细胞融合疫苗对食管癌移植瘤生长的抑制作用  

Dendritic cell-tumor cell fusion vaccine prevents growth of subcutaneous transplanted esophageal carcinomas

作　　者：邓勇军[1,2,3] 张兰军[1,2] 苏晓东[1,2] 张冬坤[1,2] 戎铁华[1,2] 王其京[1] 夏建川[1] 

机构地区：[1]华南肿瘤学国家重点实验室,广东广州510060 [2]中山大学肿瘤防治中心胸科,广东广州510060 [3]昆明医学院第一附属医院胸外科

出　　处：《癌症》2009年第10期1067-1071,共5页Chinese Journal of Cancer

基　　金：广州市科技攻关引导项目(No.2004Z3-E0311)~~

摘　　要：背景与目的:我们前期的研究表明树突细胞/肿瘤细胞融合疫苗能有效诱导体外抗食管癌细胞的特异性免疫应答,在此基础上,本研究进一步探讨树突细胞/肿瘤细胞融合疫苗诱导抗原特异性细胞毒性T淋巴细胞(cytotoxic T lymphocytes,CTLs)对食管癌细胞株EC-109裸鼠皮下移植瘤的体内抑瘤作用及对肿瘤细胞增殖和凋亡的影响。方法:①采用聚乙二醇法制备树突细胞/肿瘤细胞融合疫苗并诱导抗原特异性CTLs产生。②建立食管癌EC-109细胞裸鼠皮下移植瘤模型,将成功荷瘤的裸鼠随机分为3组,分别以融合细胞激活的CTLs(A组,n=11)、未激活的T淋巴细胞(B组,n=11)以及RMPI-1640培养液(C组,n=11)作为效应细胞进行瘤内注射,每周一次。③4周后处死动物剥离肿瘤组织,测量并比较各组肿瘤组织的平均体积及平均湿重,计算抑瘤率。④免疫组化SP法检测3组裸鼠移植瘤增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)的表达。⑤流式细胞仪检测3组肿瘤细胞周期及细胞凋亡率,比较各组肿瘤细胞的S期细胞平均百分比(S-phase fraction,SPF)以及细胞平均凋亡率。结果:①A组肿瘤组织的平均体积显著小于B组和C组,差异有统计学意义[(881.45±31.14)mm3vs.(1493.37±51.67)mm3,(2065.77±87.55)mm3,F=950.67,P=0.00],其平均湿重亦显著小于B组和C组,差异有统计学意义[(0.88±0.04)gvs.(1.38±0.07)g,(2.04±0.11)g,F=1335.90,P=0.00];A组抑瘤率明显高于B组,差异有统计学意义(56.86%vs.32.35%,F=1218.08,P=0.001)。②A组肿瘤的平均PCNA-LI明显低于B组和C组,差异有统计学意义[(26.83±0.95)%vs.(51.82±1.51)%,(68.93±2.40)%,F=1528.39,P=0.000]。③A组肿瘤细胞平均SPF值明显低于B、C两组,差异有统计学意义[(12.46±0.36)%vs.(29.39±0.96)%,(42.25±1.43)%,F=2188.05,P=0.001]。④A组肿瘤细胞平均凋亡率明显高于B、C两组,差异有统计学意义[(38.03±1.21)%vs.(17.75±0.56)%,(6.59±0.22)%,F=4565.51,P=0.001]。结论:本�Background and Objective： Our previous studies have shown that dendritic cell （DC）-tumor cell fusion vaccine can induce specific antitumor response against esophageal carcinoma cells. This study was to investigate the inhibitory effect of intratumor injection of the antigen-specific cytotoxic T lymphocytes （CTLs） induced by DC-tumor cell fusion vaccine against subcutaneously transplanted esophageal carcinoma cells in nude mice, and to analyze the influence of DC/tumor cell fusion vaccine on proliferation and apoptosis of esophageal carcinoma cells. Methods： Fusion cell vaccine of mature DOs with EC109 cells were generated by the polyethylene glycol （PEG） protocol and the antigen-specific CTLs were induced. The models of transplanted human esophageal carcinoma in nude mouse were established using EC-109 cell line. Thirty-three nude mice with subcutaneous tumors were randomly divided into three groups. Subcutaneous tumors of group A （n= 11 ）, group B （n=11） and group C （n=11） were intratumorally injected with the CTLs induced by DO/tumor fusion vaccine, T lymphocytes and RPMI 1640 medium respectively once a week. After four weeks of intratumor injection, the nude mice were killed and the nodules were anatomized. The mean volume and weight of tumors of each group were measured, and the tumor inhibitory rates of the Group A and the Group B were calculated and compared. The expression of proliferating cell nuclear antigen （PCNA） was detected by immunohistochemistry （S-P method）. The mean PCNA-label index （LI） of three groups was compared. The cell cycle and cell apoptosis of the xenograft tumor cells were analyzed by flow cytometry. The mean S- phase fraction （SPF） and the mean rate of cell apoptosis of three groups was compared respectively. Results： Both the mean volume and the mean weight of xenograft tumors in group A （881.45±31.14 mm^3 and 0.88±0.04 g） were significantly smaller than those of group B （1493.37±51.67 mm^3 and 1.38±0.07 g） and group C （2065

关 键 词：食管肿瘤 树突细胞 融合疫苗 移植瘤 细胞毒性T细胞 

分 类 号：R735.1[医药卫生—肿瘤]