重组质粒pEGFP—GPC3的抗小鼠肝癌研究  

Study of anti-mouse hepatoceHular carcinoma effects of recombinant eukaryotic plasmid pEGFD-GPC3

作　　者：吕庆杰[1] 刘剑勇[1] 赵荫农[1] 刘春辉[1] 张志明[1] 冯钟煦[1] 崔英[1] 梁新强[1] 唐凯[1] 

机构地区：[1]广西医科大学附属肿瘤医院肝胆外科,南宁530000

出　　处：《国际免疫学杂志》2013年第2期142-148,共7页International Journal of Immunology

基　　金：广西医疗卫生重点科研课题（重200611）;广西科学研究与技术开发计划项目（桂科攻0719006-2-5）;广西科学基金项目（桂科自0728196）

摘　　要：目的探讨pEGFP—GPC3转染小鼠骨髓来源树突状细胞（DC）是否能诱导小鼠产生特异性抗肝癌免疫以达到抗肝癌目的。方法建立荷瘤小鼠模型，随机分A、B、C三组，分别予腹腔注射pEGFP—GPC3转染的DC、pEGFP—N1转染的DC及生理盐水，提取各组小鼠脾淋巴细胞（脾LC）检测细胞毒性T淋巴细胞（CTL）活性，观察瘤体内淋巴细胞浸润及肿瘤细胞坏死情况并计算抑瘤率。结果A组小鼠脾LC对小鼠H22肝癌细胞（H22细胞）的杀伤率相比差异明显高于其对小鼠C127乳腺癌细胞（C127细胞）的杀伤率（t=17．365，P〈0．01）；B组小鼠脾LC对H22细胞及C127细胞的杀伤率无统计学意义（t=0．654，P〉0．05）；C组小鼠脾Lc对H22细胞及C127细胞的杀伤率无统计学意义（t=1．291，P〉0．05）；A、B、C组对H22细胞杀伤率分别比较，A〉B〉C（F=587．658，P〈0．01）。A组瘤体石蜡病理切片淋巴细胞浸润情况明显高于B组和C组（，=1．075，P〈0．01）。以C组为对照组，A、B组抑瘤率比较：A组〉B组。结论重组质粒pEGFP—GPC3转染的DC能显著提高小鼠机体对H22肝癌细胞的特异性免疫。Objective To investigate the machenism of mouse bone marrow-derived dendritic cells （DC） transfected with recombinant eukaryotic plasmid pEGFP-GPC3 inducing specific antitumor responses against hepatocellular carcinoma in mice. Methods Tumor-bearing mice were divided randomly into three gourps （A,B,C） for the injection of DC transfected with pEGFP-GPC3, DC transfected with pEGFP-N1 and physiological saline, respectively. CTL activities of splenic lymphocytes obtained from each group were tested. Tumor infiltrating lymphocytes and the level of tumor necrosis were further observed to assess the tumor inhibition rate. Results The cytotoxic activity of spleen lymphocytes on mice H22 hepatocellular carcinoma cells （H22 cells） was found significantly higher than that on mice C127 breast cancer cells （C127 cells） in the pEGFP-GPC3 gourp （ t = 17. 365, P 〈 0.01 ）. No statistically difference between the cytotoxic activity of spleen lymphocytes on H22 cells and on C127 cells was found in the other two groups （t = 0. 654, P 〉 0. 05; t = 1. 291, P 〉0.05, respectively）. The cytotoxic activity of spleen lymphocytes on H22 cells was highest in the pEGFP-GPC3 group, while lowest in the physiological saline group （ F = 587. 658, P 〈 0.01 ）. The tumor lymphocytes infiltration rate was the highest in the pEGFP-GPC3 gourp （X2 = 1. 075E, P 〈 0. 01 ）. while the lowest in the the physiological saline group. In additon, the tumor inhibition rate of the pEGFP-GPC3 group was higher than the pEGFP-N1 group. Conclusion DC transfected with recombinant eukaryotic plasmid pEGFP- GPC3 may induce specific immune responses against H22 ceils.

关 键 词：GPC3 重组质粒 pEGFP—GPC3 树突状细胞 

分 类 号：R735.7[医药卫生—肿瘤]