卵巢癌细胞B7-H4基因高表达抑制CTL细胞杀伤作用的研究  

B7-H4 expression of ovarian cancer cell inhibite the cytoxicity of CTL cells

作　　者：成磊[1] 姜洁[1] 李少儒[1] 魏双燕[2] 南芳芳[2] 

机构地区：[1]山东大学齐鲁医院妇产科,济南250012 [2]滨州医学院附属医院妇产科

出　　处：《现代妇产科进展》2009年第4期253-256,共4页Progress in Obstetrics and Gynecology

基　　金：国家自然科学基金(No:30672233);山东省自然科学基金(No:Y2006C64)

摘　　要：目的:探讨卵巢癌细胞B7-H4基因高表达对靶向诱导的CTL细胞体外杀伤的影响。方法:RT-PCR法扩增编码基因,构建真核表达载体PEGFP-N1/B7-H4,脂质体介导将重组质粒PEGFP-N1/B7-H4导入SKOV3细胞中,筛选建立高表达细胞株。细胞免疫组织化检学检测B7-H4蛋白的表达。利用免疫磁珠分离法(MACS)分离纯化脐血CD34+细胞并在体外诱导分化为DC,用反复冻融法从卵巢癌细胞SKOV3中提取可溶性冻融抗原并负载DC诱导生成CTL。MTT法检测经抗原负载后激活的CTL细胞对SKOV3/B7-H4、SKOV3/neo和SKOV3细胞的杀伤作用。结果:成功构建了人B7-H4真核表达载体,SK-OV3/B7-H4细胞稳定表达B7-H4蛋白。CTL细胞对SKOV3/B7-H4细胞杀伤作用明显低于阴性对照组(20.12%±3.14%vs36.34%±4.53%,P<0.05)。结论:卵巢癌细胞B7-H4基因表达可抑制靶向诱导的CTL细胞毒性,可能与肿瘤细胞免疫逃逸有着密切的关系。Objective：To explore the effect of B7-H4 expression of ovarian cancer cell on cytoxicity of CTL cells. Methods：The PEGFP-N1/B7-H4 vector was constructed,and transfected into B7-H4-negative human epithelial ovarian cancer SKOV3 cells. After isolated and purified from umbilical cord blood by a higher gradient magnetic cell sorting system （MASC） , CD34^＋ cells were induced to differentiate into DC by cocktail cytokines. Freeze-thaw antigens were extracted from SKOV3 cells by freeze-thaw method, and then loaded on DC. The cytotoxicity of CTL activated by DC loading with freeze-thaw ovarian cancer antigen of SKOV3/B7-H4 SKOV3/neo and SKOV3 was evaluated by 3- （4,5- dimethyhhiazol-2-yl） -2,5-diphenyltetrazolium bromide （MTT） assay. Result：The eukaryotic expression vector of PEGFP-N1/B7-H4 was constructed and verified successfully. The B7-H4 expression was positive in SKOV3/B7-H4 cells through immunohistochemistry analyze, but was not in control cells. The killing rate of CTL on SKOV3/B7-H4 was significantly lower than that on SKOV3/neo （ 20. 12% ±3. 14% vs 36. 34% ±4.53% ）. Conclusion： B7-H4 expression can inhibite the cytoxicity of CTL. It may play a more pivotal role in tumor immune escape.

关 键 词：B7-H4 卵巢肿瘤 质粒构建 CTL 

分 类 号：R737.33[医药卫生—肿瘤]