人源化肿瘤血管移植瘤模型的建立及应用  

Establishment and Application of Engrafted Tumor Models with Humanized Tumor Blood Vessel

作　　者：冉宇靓[1] 钟星[1] 胡海[1] 遇珑[1] 娄晋宁[2] 杨治华[1] 

机构地区：[1]中国医学科学院中国协和医科大学肿瘤医院/肿瘤研究所细胞生物及分子生物学室,北京100021 [2]中日友好医院临床医学研究所,北京100029

出　　处：《癌症》2006年第11期1323-1328,共6页Chinese Journal of Cancer

基　　金：国家自然科学基金重点项目(No.30230150);国家重点基础研究发展规划"973"项目(No.2002CB513106)~~

摘　　要：背景与目的:目前在肿瘤内皮细胞基因功能和靶向血管治疗肿瘤的研究中仍缺乏适宜的动物模型。本研究拟建立一种新的小鼠人源化肿瘤血管移植瘤模型。方法:将人肝窦微血管内皮细胞系(humanliversinusoidendothelialcells,HLSEC)分别与人肝癌细胞系BEL7402、人结肠癌细胞系LS174T、人食管癌细胞系NEC按不同比例混合共接种NOD/SCID小鼠或BALB/c裸小鼠,以单独接种肿瘤细胞的小鼠作为对照组,观察小鼠人移植瘤生长的情况。采用绿色荧光蛋白基因(greenfluorescentprotein,GFP)转染HLSEC,结合荧光显微镜检方法观察HLSEC在共接种移植瘤中的存活及血管形成的情况。免疫组化法检测肿瘤内微血管密度(microvesseldensity,MVD)。用抗人肝癌内皮细胞单抗2B6处理人肝癌移植瘤共接种模型,观察2B6对肿瘤生长的影响。结果:HLSEC与BEL7402细胞共接种NOD/SCID小鼠时,共接种组肿瘤生长速度显著加快,平均瘤重可达肿瘤单独接种组的5.1倍。在GFP表达阳性的HLSEC与BEL7402共接种的移植瘤冰冻切片中可见HLSEC的存在,并已形成瘤内新生血管。免疫组化检测发现共接种组移植瘤中的总MVD较肿瘤细胞单独接种组增加85.7%,采用抗人vWF多抗检测结果显示共接种组人源微血管平均MVD可达10.28～29.28,约占总血管数量的41%～65%。进一步将HLSEC分别与NEC、BEL7402及LS174T细胞共接种于BALB/c裸小鼠时,共接种组的瘤重是单独接种组的3.3～6.0倍。2B6单抗能使共接种人肝癌移植瘤中人源肿瘤血管密度减少65.1%,抑制肿瘤瘤重达71.8%。结论:HLSEC与人肿瘤细胞共接种小鼠后,能在移植瘤中存活、增殖,形成大量人源化的肿瘤新生血管,并在促进肿瘤快速生长中起重要作用。该小鼠人源化肿瘤血管移植瘤模型可为肿瘤内皮细胞相关基因及靶向治疗剂的研究提供一个新的有价值的工具。BACKGROUND ＆ OBJECTIVE. Animal models are indispensable in the studies of tumor endothelial genes and anti-angiogenic therapy. This study was to build a new engrafted tumor model with humanized blood vessels. METHODS： Human liver sinusoid endothelial cells （HLSECs） were mixed respectively with human liver cancer cell line BEL7402, human colon cancer cell line LS174T, and human esophageal cancer cell line NEC, and then inoculated into NOD/SCID mice or BALB/c nude mice. The mice inoculated with only tumor cells were used as controls. Tumor growth was observed. Green fluorescent protein （GFP）-Iabeled HLSECs were observed under fluorescent microscope to detect their survival and tube formation. Microvessel density （MVD） was analyzed by immunohistochemistry. The tumor-bearing mice were treated by anti-HLSEC monoclonal antibody 2B6 to observe its effect on tumor growth. RESULTS, Tumor growth was significantly enhanced by the co-inoculation of HLSECs with BEL7402 cells in NOD/SCID mice; tumor weight was increased by 5.1 folds as compared with that of control. GFP-labeled vessels could easily be observed in the tumors from co-inoculation of HLSECs with BEL7402 cells. Total MVD was increased by 85.7% of control. The humanized MVD was about 10.28-29.28, which was 41%-65% of total MVD. Furthermore, the co- inoculation of HLSECs with NECs, BEL7402, or LS174T cells in nude mice led to 3.3-6.0 folds increase of xenograft weight as compared with control. When treated with 2B6 antibody, humanized MVD was decreased by 65.1% in the engrafted tumors and the tumor weight lost 71.8%. CONCLUSIONS： When co-inoculate with human tumor cell lines into mice, HLSECs could survive, proliferate, and contribute to tumor angiogenesis, which may enhance tumor growth. The engrafted tumor in vivo model with humanized vessels can be widely used in the research of functional genes in tumor angiogenesis and anti-angiogenic therapies.

关 键 词：微血管内皮细胞 人源化肿瘤血管 移植瘤模型 微血管密度 小鼠 

分 类 号：R73-3[医药卫生—肿瘤]