Abstract
To explore a new model of in-situ xenograft lymphangiogenesis of human colonic adenocarcinomas in nude mice. On the basis of establishing subcutaneous xenograft lymphangiogenesis model of human colonic adenocarcinoms, in-situ xenografts were established through the in situ growth of the HT-29 human colonic adenocarcinoma cell line in nude mice. The numbers of lymphangiogenic microvessels, the expression of lymphatic endothelial cell markers lymphatic vessel endothelial hyaloronic acid receptor-1 (LYVE-1), D2-40 and the 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. 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, identical with the morphological characteristics of lymphatic vessels and capillaries. Expression of LYVE-1 and D2-40 proteins and mRNAs were significantly higher in xenografts in-situ than in 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), in conformity with the signal regulation of the VEGF-C,-D/VEGFR-3 axis of tumor lymphangiogenesis. In-situ xenografts of a human colonic adenocarcinoma cell line demonstrate tumor lymphangiogenesis. This novel in-situ animal model should be useful for further studying mechanisms of lymph node metastasis, drug intervention and anti-metastasis therapy in colorectal cancer.
Highlights
Colorectal adenocarcinoma is the most common malignant tumor via the lymphatic metastasis in human
In order to study lymphatic metastasis in colorectal cancer and antilymphangiogenic treatment, it is imperative to establish a line of lymphangiogenic animal models to research human colorectal cancer (Fidler, 2003)
Scholars have reported using an orthotopic model of human colorectal cancer transplantation, in order to improve the human colorectal cancer regional lymph node metastasis rate, but no tumor lymphangiogenesis in animal models have been reported
Summary
Colorectal adenocarcinoma is the most common malignant tumor via the lymphatic metastasis in human. In order to study lymphatic metastasis in colorectal cancer and antilymphangiogenic treatment, it is imperative to establish a line of lymphangiogenic animal models to research human colorectal cancer (Fidler, 2003). Many previous studies take use of xenograft human colorectal carcinoma in nude mice as a common animal model Such a model of subcutaneous tumors DUH XVXDOO\ FRYHUHG E\ ÀEHU OHDGLQJ WR OHVV O\PSKDWLF and hematogenous metastasis, so it is not exactly the performance of biological characteristics in colorectallymphatic or hematogenous metastasis. Our experiment built an orthotopic model of human colorectal cancer transplantation ,on the basis of colorectal cancer subcutaneous xenograft model in nude mice, to explore the possibility of the establishment of human colon adenocarcinoma lymphangiogenesis model in nude mouse, in order to provide experimental models for study of human colorectal cancer lymphatic metastasis mechanisms and treatment
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