Ran Yu-Liang, Zhong Xing, Hu Hai, Yu Long, Lou Jin-Ning, Yang Zhi-Hua
Department of Cellular and Molecular Biology, Cancer Institute/Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, 100021, P. R. China.
Ai Zheng. 2006 Nov;25(11):1323-8.
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.
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)-labeled 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.
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%.
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.
动物模型在肿瘤内皮基因及抗血管生成治疗研究中不可或缺。本研究旨在构建一种具有人源化血管的移植瘤新模型。
将人肝窦内皮细胞（HLSECs）分别与人肝癌细胞系BEL7402、人结肠癌细胞系LS174T和人食管癌细胞系NEC混合，然后接种到NOD/SCID小鼠或BALB/c裸鼠体内。仅接种肿瘤细胞的小鼠作为对照。观察肿瘤生长情况。在荧光显微镜下观察绿色荧光蛋白（GFP）标记的HLSECs，以检测其存活及成管情况。通过免疫组织化学分析微血管密度（MVD）。用抗HLSEC单克隆抗体2B6处理荷瘤小鼠，观察其对肿瘤生长的影响。
在NOD/SCID小鼠中，HLSECs与BEL7402细胞共接种显著促进了肿瘤生长；肿瘤重量比对照组增加了5.1倍。在HLSECs与BEL7402细胞共接种的肿瘤中，可轻易观察到GFP标记的血管。总MVD比对照组增加了85.7%。人源化MVD约为10.28 - 29.28，占总MVD的41% - 65%。此外，在裸鼠中，HLSECs与NECs、BEL7402或LS174T细胞共接种导致异种移植瘤重量比对照组增加了3.3 - 6.0倍。用2B6抗体处理后，移植瘤中的人源化MVD降低了65.1%，肿瘤重量减轻了71.8%。
当与人类肿瘤细胞系共接种到小鼠体内时，HLSECs能够存活、增殖并促进肿瘤血管生成，这可能会促进肿瘤生长。具有人源化血管的体内移植瘤模型可广泛应用于肿瘤血管生成功能基因及抗血管生成治疗的研究。
