新城疫病毒对Hep-2肿瘤细胞的抑制效应  

Antitumor research on human laryngeal carcinoma cell line Hep-2 with newcastle disease virus

作　　者：管国芳[1] 金春顺[1] 金宁一[2] 米志强[2] 李霄[2] 连海[2] 孙丽丽[1] 文连姬[1] 

机构地区：[1]吉林大学白求恩医学部第二临床医院耳鼻咽喉科,吉林长春130041 [2]军事医学科学院全军基因工程重点实验室,吉林长春130062

出　　处：《中国耳鼻咽喉头颈外科》2005年第8期487-490,共4页Chinese Archives of Otolaryngology-Head and Neck Surgery

基　　金："973"规划(G199011902);自然科学基金重大项目资助(39893290-4-3)

摘　　要：目的探讨新城疫病毒(newcastlediseasevirus,NDV)对Hep-2肿瘤细胞的杀伤效应及其免疫刺激作用。方法采用考马斯亮兰染色法观察NDV感染对Hep-2肿瘤细胞细胞骨架的影响。运用甲基噻唑基四唑(methylthiazolyltetrazolium,MTT)染色法检测NDV感染致Hep-2肿瘤细胞死亡的杀伤率。通过免疫荧光结合流式细胞仪(flowcytometer,FCM)检测Hep-2肿瘤细胞表面主要组织相容性复合体Ⅰ类分子(majorhistocompabilitycomplexclassImolecules,MHC-I)的表达情况。另外,分离人外周血淋巴细胞(peripheralbloodmononuclearcells,PBMC)作为效应细胞,以NDV感染的Hep-2肿瘤细胞作为靶细胞,同时设空白细胞对照,通过特异性细胞毒T淋巴细胞实验,探讨NDV所介导的抑瘤作用。结果NDV可致Hep-2肿瘤细胞的细胞骨架发生改变,对体外培养Hep-2肿瘤细胞的杀伤率达77.7%,可上调其表面MHC-I类分子的表达,并且能够刺激增强人外周血淋巴细胞中肿瘤特异性细胞毒性T淋巴细胞(cytotoxicTlymphocyte,CTL)活性,杀伤率为23.54%。结论NDV可有效杀伤Hep-2肿瘤细胞,并刺激产生特异性抗肿瘤免疫反应。OBJECTIVE To study the immuno-stimulatory and anti-tumor effects of newcastle disease virus （NDV） on Hep-2 cell line. METHODS Coomassie Brilliant Blue R250 was used to observe the effect of NDV on cytoskeleton of Hep-2 cells. Methyl thiazolyl tetrazolium（MTT） method was used to detect anti-tumor effect of NDV on Hep-2 cells. The major histocompability complex class Ⅰ molecules （MHC- Ⅰ） molecule expression was detected by FCM through immunofluorescence. Human peripheral blood lymphocytes were separated and used as effector cells. Hep-2 cells infected with NDV were used as target cells. Anti-tumor mechanism was evaluated with specific cytotoxic T lymphocyte （CTL） assay. RESULTS NDV infection resulted in cytoskeleton changes of Hep-2 cells. The tumor suppression rate of NDV was 77.7 % and the specific CTL activity was 23.45 %. Furthermore, NDV infection can up-regulate the MHC-Ⅰ expression in Hep-2 cells. CONCLUSION NDV can kill Hep-2 cells efficiently in vitro and have notable immunostimulatory ability.

关 键 词：新城疫病毒 喉肿瘤 细胞 免疫法 肿瘤细胞死亡 HEP-2 特异性细胞毒性T淋巴细胞 抑制效应 人外周血淋巴细胞 MHC-1类分子 

分 类 号：R73-36[医药卫生—肿瘤]