基础医学与临床 ›› 2010, Vol. 30 ›› Issue (5): 524-529.

• 研究论文 • 上一篇    下一篇

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

刘虹麟 吴练秋 叶丽亚 娄晋宁 张文健   

  1. 北京协和医学院研究生院 北京协和医学院研究生院 中日友好医院临床医学研究所 中日友好医院临床医学研究所 中日友好医院临床医学研究所
  • 收稿日期:2009-12-28 修回日期:2010-01-08 出版日期:2010-05-05 发布日期:2010-05-05
  • 通讯作者: 张文健

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

Hong-lin LIU, Lian-qiu WU, Li-ya YE, Jin-ning LOU, Wen-jian ZHANG   

  1. Granduate School, PUMC Institute of Clinical Medical Sciences, China-Japan Friendship Hospital Institute of Clinical Medical Sciences, China-Japan Friendship Hospital
  • Received:2009-12-28 Revised:2010-01-08 Online:2010-05-05 Published:2010-05-05
  • Contact: Wen-jian ZHANG

摘要: 目的 比较淋巴管内皮细胞对乳腺癌和骨肉瘤生长和转移的影响,分析淋巴管生成对肿瘤生长和转移影响的相关机制。方法 应用抗人VEGFR-3抗体包被的磁珠从人淋巴结中分离纯化人淋巴管内皮细胞(HLyEC)。将人乳腺癌细胞株MDA-MB-435、人骨肉瘤细胞株MG-63单独或分别与HLyEC共同接种于裸鼠胸部皮下,比较肿瘤生长的差别。应用伊文氏蓝染料瘤内注射显示肿瘤周围的淋巴管;应用人PDPN和小鼠PDPN的免疫组织化学染色评价肿瘤组织中的淋巴管密度和来源,计数肺表面肿瘤的转移克隆数。用MTT方法分析乳腺癌和骨肉瘤细胞条件培养基对淋巴管内皮细胞增殖的影响。结果 共接种HLyEC促进乳腺癌细胞的生长,但对骨肉瘤细胞的生长没有影响。共接种淋巴管内皮的乳腺癌组织中淋巴管密度明显增加,与单纯接种组有显著差异,肿瘤内存在人PDPN阳性和鼠PDPN阳性的淋巴管;而共接种HLyEC对骨肉瘤内淋巴管密度无影响,瘤组织内未见人或小鼠PDPN阳性淋巴管。共接种HLyEC的乳腺癌组可见明显肺表面转移克隆,而共接种HLyEC的骨肉瘤组和两个单纯接种肿瘤组均未见肺表面转移灶。MTT分析结果显示:乳腺癌细胞的条件培养基明显促进淋巴管内皮细胞的增殖,而骨肉瘤细胞条件培养基对淋巴管内皮细胞增殖没有影响。结论 乳腺癌细胞可能通过活跃的淋巴管生成加速肿瘤的生长和转移。

关键词: 淋巴管内皮细胞, 乳腺癌, 骨肉瘤, 淋巴管生成, 转移

Abstract: Objective To compare growth and metastasis of breast cancer cell and osteosarcoma cell influenced by co-inoculating human lymphatic endothelial cell (HLyEC), and to analyze its mechanisms. Methods HLyEC was 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 HLyEC subcutaneously into nude mice. Weight of tumors were detected; the peri-tumor lymphatic vessel density was shown by Evans blue, the intra-tumor lymphatic vessel density and origin of intra-tumor lymphatic endothelial cell were evaluated by immunohistochemistry for human and mouse PDPN. And the metastasis clones on lungs was counted. The effect of MDA-MB-435 and MG-63 condition medium on proliferation of HLyEC was evaluated by MTT assay. Results The growth of MDA-MB-435 but not MG-63 was promoted by co-inoculating HLyEC with much higher weight of tumor. The peri-tumor and intra-tumor lymphatic vessel density of MDA-MB-435 were increased by co-inoculating HLyEC, both hPDPN- and mPDPN-positive lymphatic vessels were found in MDA-MB-435 co-inoculating with HLyEC group. But co-inoculating HLyEC has no effect on lymphatic vessels density of MG-63. Neither hPDPN- nor mPDPN-positive lymphatic vessels were found in MG-63 tumor. And many metastasis clones were visible on lungs only at MDA-MB-435 co-inoculating with HLyEC group. The result of MTT assay showed that the proliferation of HLyEC was increased by condition medium of MDA-MB-435 but not MG-63. Conclusion Breast cancer has active lymphangiogenesis which might contribute to its growth and metastasis.

Key words: lymphatic endothelium, breast cancer, osteosarcoma, lymphaniogenesis, metastasis