共接种淋巴管内皮细胞对乳腺癌细胞和骨肉瘤细胞在裸鼠体内生长和转移的影响

The effect of co-inoculating human lymphatic endothelial cells on growth and metastasis of breast cancer cell and of osteosarcoma cell in nude mice

目的分析淋巴管生成对肿瘤生长和转移的影响。方法从人淋巴结中分离纯化淋巴管内皮细胞(HLyECs),将人乳腺癌细胞系和人骨肉瘤细胞系分别单独或与HLyECs共同接种于裸鼠皮下,比较肿瘤生长和肺转移的差别。用伊文氏蓝显示瘤周淋巴管;用人和小鼠PDPN的免疫组织化学显示肿瘤组织中的淋巴管。用MTT法分析淋巴管内皮细胞的增殖。结果与单纯接种组相比,共接种HLyECs促进乳腺癌细胞的生长和转移,瘤周和瘤内淋巴管密度增加(P<0.01),存在人和鼠PDPN阳性的淋巴管;而共接种HLyECs对骨肉瘤的生长和转移无影响,未见瘤内和瘤周淋巴管。与骨肉瘤细胞相比,乳腺癌细胞的条件培养基明显促进淋巴管内皮细胞增殖(P<0.01)。结论共接种淋巴管内皮细胞可以促进乳腺癌细胞的生长及癌组织中淋巴管生成。

Objective To investigate the effects of lymphangiogenesis on cancer growth and metastasis.MethodsHuman lymphatic endothelial cells（HLyECs） were isolated and purified from human lymph node by magnetic beads coated with antibody against human VEGFR3.Human breast cancer cell line（MDA-MB-435） or human osteosarcoma cell line（MG-63） was inoculated alone or co-inoculated with HLyECs subcutaneously into nude mice.The weight of tumor and lung surface metastasis were detected;peri-tumor lymphatic vessels were shown by Evans blue,intra-tumor lymphatic vessels were shown by immunohistochemistry for human and mouse PDPN.Conditioned medium of tumor on proliferation of HLyECs was evaluated by MTT assay.Results Compared with inoculating tumor cells alone,the growth and metastasis of MDA-MB-435 were promoted by co-inoculating HLyECs.The peri-tumoral and intra-tumoral lymphatic vessel density of MDA-MB-435 were increased by co-inoculating HLyECs.Both hPDPN-and mPDPN-positive lymphatic vessels were found.But co-inoculating HLyECs had no effect on lymphatic vessels density of MG-63.Neither intra-nor peri-tumor lymphatic vessels were found.The proliferation of HLyECs was increased by conditioned medium of MDA-MB-435 but not by MG-63.Conclusion Growth and lym-phangiogenesis of breast cancer can be enhanced by co-inoculating lymphatic endothelial cells.