一种新的人结肠腺癌淋巴管生成裸鼠动物模型

A new model of in-situ xenograft lymphangiogennesis of human colonic adenocarcinomas in nude mice

目的探索一种新的人结肠腺癌淋巴管生成裸鼠动物模型。方法在建立人结肠腺癌皮下移植瘤基础上构建人结肠腺癌裸鼠原位移植瘤裸鼠模型,通过淋巴管内皮细胞标记LYVE-1、D2-40和淋巴管生成因子VEGF-C、VEGF-D及其受体VEGFR-3的免疫组化、Western blot和荧光定量RT-PCR检测以鉴定是否为肿瘤淋巴管生成。结果与结肠阴性对照组即无瘤组比较,结肠原位种植瘤中呈LYVE-1、D2-40免疫组化染色强阳性的管道管壁较薄、管腔大而不规则或呈塌陷状,微淋巴管密度(LMVD)高,符合毛细淋巴管形态学特征;而且LYVE-1、D2-40蛋白和mRNA表达明显升高(P<0.01);同时,结肠原位种植瘤的VEGF-C,VEGF-D和VEGFR-3蛋白和mRNA表达亦明显高于无瘤组(P<0.01),支持肿瘤淋巴管生成的VEGF-C,-D/VEGFR-3信号调控机制。结论裸鼠结肠癌原位种植瘤存在淋巴管生成。该肿瘤淋巴管生成模型可作为一种新的人结肠腺癌淋巴管生成动物模型,为深入研究结直肠癌淋巴管生成和转移机制、药物干预及抗淋巴转移治疗提供参考。

Objective To explore a new model of in-situ xenograft lymphangiogennesis of human colonic adenocarcinomas in nude mice.Method On the basis of establishing subcutaneous xenograft lymphangiogenesis model of human colonic adenocarcinoms,in-situ xenograft lymphangiogenic models of human colonic adenocarcinomas were established through the in situ growth of human colonic adenocarcinoma cell line HT-29 in nude mice.The number of lymphangiogenetic microvessels,the expression of lymphatic endothelial cell markers lymphatic vessel endothelial hyaloronic acid receptor-1 (LYVE-1),D2-40 and lymphatic endothelial growth factors vascular endothelial growth factor-C (VEGF-C),-D (VEGF-D) and receptor-3 (VEGFR-3) were compared by immunohistochemical staining,Western bolt and quantitative RT-PCR in xenograft in-situ models.Results Some microlymphatics with thin walls,large and irregular or collapsed cavities and increased LMVD,with strong positive of LYVE-1,D2-40 in immunohistochemistry,were observed,and identical with the morphological characteristic of the lymphatic vessels and capillaries.And,the expression of LYVE-1 and D2-40 proteins and mRNAs were significantly higher in xenografts in-situ than those of the negative control group (both P<0.01).Moreover,the expression of VEGF-C,VEGF-D and VEGFR-3 proteins and mRNAs were significantly higher in xenografts in-situ (both P<0.01),that was in conformity with the signal regulation of VEGF-C,-D/VEGFR-3 axis of tumor lymphangiogennesis.Conclusions In situ xenografts of human colonic adenocarcinomas there have been tumor lymphangiogenesis.The in-situ model of xenograft lymphangiogenesis may act as an ideal,referential,new animal model for further studying of lymph node metastasis mechanism,drug intervention and anti-metastasis therapy in colorectal cancer.