HA纳米载体转GM-CSF基因制备自体肿瘤疫苗治疗肝癌的实验研究

Autologous Tumor Vaccine of the Transfected GM-CSF Gene Mediated by Nano-HA Vector Inhibits Xenografes Growth in Human Hepatocellular Carcinoma PBL-SCID Chimeric Model

目的:观察自体肿瘤疫苗对人肝癌PBL-SCID嵌合模型的治疗效果。方法:SCID小鼠20只腹腔内注射健康志愿者外周血淋巴细胞(2×10~6/mL)0.5 mL,同时背部皮下接种人肝癌HepG2细胞(1×10~7/mL)0.2mL。当皮下移植瘤体积长至100mm^3时,随机分4组:Ⅰ组,生理盐水组;Ⅱ组,^(60)Co照射的HepG2细胞组;Ⅲ组,转染GM-CSF基因的HepG2细胞组;Ⅳ组,^(60)Co照射的转染GM-CSF基因的HepG2细胞组即自体肿瘤疫苗组,每组5只。结果:自体肿瘤疫苗组6周存活率(100%)高于生理盐水组(60%)和单纯^(60)Co照射的HepG2细胞组(40%);自体肿瘤疫苗腹腔注射抑制皮下移植瘤生长,其肿瘤生长抑制率为89%;免疫组化检测发现人淋巴细胞分布于小鼠移植瘤组织中。病理切片见移植瘤细胞变性和死亡。结论:HA纳米载体转GM-CSF基因制备的自体肿瘤疫苗具有抑制人肝癌PBL-SCID嵌合模型的皮下移植瘤生长作用,并诱导有效的抗肿瘤免疫反应。

Objective：To observe the therapeutic effect of autologous tumor vaccine on cherimic huPBL-SCID-HCC model in SCID mice. Methods： Health human peripheral blood lymphocytes were isolated from peripheral blood of healthy donors and intraperitoneal injected into SCID mice at 1 × 10^6/mouse. Sinlultaneously,hepatocellular carcinoma cells were intracutaneously injected into SCID mouse at 2 × 10^6/mouse. When the xenografts to a 100mm3 , mice is randomly divided into 4 groups ： Group 1, normal saline group ; Group 2, HepG2 cells after ^60Co irradiation group ; Group 3, HepG2 cells transfected with GM-CSF Gene group ; Group 4, HepG2 cells transfected with GM-CSF Gene after ^6oCo irradiation group, That is autologous tumor vaccine group ; every group has 5 mice. Results ： The survival rate of mice at sixth week in group 4 （ 100% ） were higher than normal saline Group and HepG2 ceils after 60 Co irradiation group. Autologous tumor vaccine inhibits subcutaneous tumor xenograft growth. The inhibit rate of tumor growth were 89% ,The distribution of lymphocytes in transplanted tumor tissue can be detected by imnmnohistochemistry. Transplanted tmnor cells got degeneration and death on the pathological section. Conclusions ： Autologous tumor vaccine of GM-CSF Gene transfected by the nauo-hydroxyapatite vector has an inhibitory effect on growth of subcutaneous tumor of human hepatocellular carcinoma in human PBL-SCID chimeric model,and could induce effective anti-tumor immunity.