膜表达热休克蛋白70的肠癌瘤苗抗肿瘤治疗作用研究  

Antitumor therapeutic effect induced by intestine-carcinoma cells vaccine expressing membrane-bound heat shock protein70

作　　者：黄常新[1] 余海[2] 杨关根[1] 王青青[2] 沈芬平[2] 李达[2] 曹雪涛[2] 

机构地区：[1]杭州市第三人民医院肛肠科,310009 [2]浙江大学免疫学研究所

出　　处：《中华胃肠外科杂志》2005年第3期255-258,共4页Chinese Journal of Gastrointestinal Surgery

摘　　要：目的研制膜表达热休克蛋白(HSP)70瘤苗并研究其抗肿瘤治疗作用。方法以膜表达HSP70的真核表达载体pDisplay-mbHSP70转染肠癌细胞株CT26。用G418抗性筛选获得阳性克隆,扩增并制备瘤苗。观察瘤苗在体外对小鼠脾淋巴细胞的增殖刺激效应;在BALB/c小鼠结直肠癌模型中观察瘤苗的抗肿瘤治疗作用;并检测实验鼠的自然杀伤细胞(NK)和细胞毒性T淋巴细胞(CTL)活性。结果激光共聚焦显微镜和流式细胞术均证明,大量HSP70表达并结合在肠癌瘤苗细胞膜表面。比较对照,胞浆表达HSP70瘤苗显示出较高的鼠淋巴细胞增殖刺激效应,较强的瘤体生长抑制,并使荷瘤鼠的生存时间延长;而膜表达HSP70瘤苗比胞浆表达者有着更高的淋巴增殖刺激效应,更强的瘤体生长抑制作用,并使荷瘤鼠的生存时间进一步延长。结论HSP70表达并结合在细胞膜表面的肠癌细胞瘤苗对结直肠癌CT26荷瘤小鼠有更强的治疗作用。Objective To develop a new vaccine expressing membrane bound he at shock protein 70(mbHSP70) and further study its antitumor therapeutic effect. M ethod The pre established vector expressing mbHSP70 was transfected into CT26 c ells of colorectal cancer. After the CT26 cells were incubated with 900 μg/ml G 418,the sub clones resistant to G418 were harvested and the HSP70 positive clon es were selected by limiting dilution. The clones were amplified and inactivated ,thereby the vaccine expressing mbHSP70 was prepared. Lymphocyte proliferation s timulated by the vaccines,NK and CTL activity was observed. The antitumor effica cy of vaccine was observed in BALB/c mice model with colorectal cancer. Results The laser confocal microscopy and flow cytometry showed that there existed much HSP70 on the vaccine membrane surface. The HSP70 gene modified vaccine displaye d augmented lymphocyte proliferation and higher NK and CTL activity in vitro,and marked tumor suppression and prolonged survival time of the vaccinated mice in vivo,when compared with its counterpart. Furthermore,mbHSP70 expression vaccine elicited lymphocyte proliferation most intensively,generated the highest NK and CTL activity as well as the strongest antitumor effect,and prolonged survival t ime of the vaccinated mice. Conclusion A new vaccine expressing mbHSP70 has more potent antitumor immunity and better therapeutic efficacy than HSP70 gene modi fied vaccine did.

关 键 词：热休克蛋白70 膜表达 抗肿瘤治疗 作用研究 HSP70表达 细胞毒性T淋巴细胞 G418抗性筛选 BALB/c小鼠 激光共聚焦显微镜 治疗作用 真核表达载体 刺激效应 自然杀伤细胞 淋巴细胞增殖 生长抑制作用 结直肠癌 胞膜表面 脾淋巴细胞 

分 类 号：R735.3[医药卫生—肿瘤]