单核细胞趋化因子-1介导单核细胞治疗卵巢癌的实验研究  

Therapeutic effect of MCP-1 mediated macrophages on ovarian epithelial carcinoma

作　　者：张萍[1] 张爱荣[1] 洪淑惠[2] 孔北华[3] 吴学忠[4] 高晓林[1] 

机构地区：[1]山东大学第二医院妇产科,山东济南250033 [2]山东省千佛山医院妇产科,山东济南250014 [3]山东大学齐鲁医院妇产科,山东济南250012 [4]山东省医学科学院肿瘤免疫与基因工程重点实验室,山东济南250062

出　　处：《中国病理生理杂志》2008年第6期1114-1118,共5页Chinese Journal of Pathophysiology

基　　金：山东省医药卫生科技发展计划资助项目(No2005JW2015)

摘　　要：目的:研究单核细胞趋化蛋白-1(MCP-1)介导单核细胞对大鼠卵巢上皮癌的治疗作用并探讨其机制。方法:用脂质体介导法将质粒pLXSN/MCP-1转染包装细胞PA317,经G418筛选滴度最高的PA317病毒上清液感染大鼠卵巢癌细胞株NuTu-19,以RT-PCR和Boyden Chamber分析MCP-1的表达。分离Fischer344大鼠脾脏单核巨噬细胞,采用MTT法检测MCP-1介导单核细胞对肿瘤细胞的杀伤效应。观察基因修饰的肿瘤细胞对卵巢癌腹腔转移大鼠模型生存期的影响并采用流式细胞仪检测肿瘤组织中CD25(IL-2R)及CD44v6的表达情况。结果:本研究建立的NuTu-19/MCP-1细胞株中MCP-1稳定表达并具有较高的单核细胞趋化活性,趋化指数较对照组明显升高(P<0.05)。MCP-1体外介导单核细胞对NuTu-19/MCP-1细胞的最大杀伤率为28%,明显高于对照组(P<0.05)。基因修饰的肿瘤细胞可在一定程度上延长荷瘤动物生存期,与对照组相比差异显著(P<0.05)。FCM显示NuTu-19/MCP-1组(25.82%)细胞中CD25(IL-2R)含量显著高于对照组NuTu-19/neo(8.73%)细胞(P<0.05)。CD44v6的含量在NuTu-19/MCP-1组(6.61%)显著低于对照组(40.74%)的表达水平,差异显著(P<0.01)。结论:MCP-1可介导单核细胞抑制卵巢癌细胞NuTu-19的生长,MCP-1基因修饰的肿瘤细胞具有一定的诱导机体抗肿瘤免疫作用,MCP-1免疫基因治疗对提高卵巢癌的治疗效果具有重要意义。AIM： To investigate the therapeutic effect of MCP-1 mediated macrophages on ovarian epithelial carcinoma and its mechanisms. METHODS： Retorviral expression vectors pLXSN/MCP - 1 was transfected into the packaging cell line PA317 by lipofectin-mediated gene transfer system. The virus particles containing MCP-1 gene were collected to infect NuTu-19. RT-PCR and Boyden Chamber were used to confirm the expression of MCP-1. Rat Fischer344 spleen macrophages were isolated. MTr method was applied to investigate the tumoricidal effect of macruphages. The survival time of the intraperitoneal disseminating ovarian cancer animal model was observed, and flow cytometry method was applied to analyze the expression of CD25 or CD44v6, and then the anti - tumor mechanisms of gene modified tumor cell lines were discussed. RESULTS： Stable MCP - 1 expression in the cell line NuTu - 19/MCP - 1 possessed the chemotatic activity. The maximum killing ratio of macruphages on NuTu - 19/MCP - 1 cells was 28%. In the animal models immunized by MCP - 1 expressing cells, prolonged survival time was showed which had statistical significance compared with that in the control group （P 〈0. 05）. The expression rate of CD25 （25. 82% ） in the NuTu-19/MCP-1 cells was higher than that in NuTu - 19/neo cells （8.73%）. The expression of CD44,6 in NuTu - 19/MCP - 1 cells was significantly lower than that in control NuTu - 19/neo cells. CONCLUSION： MCP - 1 mediates macrophages and suppresses the growth of NuTu - 19. MCP - 1 gene modified tumor cells can induce anti - tumor immunity. This strategy would be used as a promising approach for the treatment of ovarian cancer.

关 键 词：炎症趋化因子类 卵巢肿瘤 基因疗法 单核细胞化学吸引蛋白质-1 

分 类 号：R737.31[医药卫生—肿瘤]