中文摘要：

该研究旨在探讨乳腺癌来源的透明质酸（hyaluronic acid,HA）分子大小变化对淋巴内皮细胞紧密连接分子ZO-1（zonula occludens-1）分布情况以及细胞增殖的影响。采用免疫组化实验检测8对良性乳腺疾病和乳腺癌患者组织HA的表达水平,利用酶联免疫吸附实验检测20对正常人与乳腺癌患者血清中透明质酸分解酶（hyaluronidase,Hyase）含量;通过免疫荧光法、荧光素钠渗透实验、MTT增殖实验和Western blot分别观察HA和寡分子透明质酸（oligosaccharides of Hyaluronic acid,o HA）作用淋巴内皮细胞后ZO-1分布、单层细胞渗透性、细胞增殖以及下游ROCK1/Rho A信号通路变化。结果表明,与良性乳腺疾病对照组比较,乳腺癌患者HA表达量明显增加（P〈0.01）;与正常人相比,乳腺癌患者血清Hyase水平显著升高（P〈0.01）,提示乳腺癌发生时,HA升高伴随其降解酶增多,HA代谢活性增加。HA增强细胞间ZO-1的连接,对ROCK1/Rho A蛋白表达水平无明显影响;相反,o HA促使淋巴内皮细胞ZO-1由胞膜向胞浆分布,导致细胞连接呈褶皱状,形成细胞间隙,淋巴管渗透性增加,上调ROCK1/Rho A蛋白表达水平,促进淋巴内皮细胞增殖（P〈0.01）。研究提示,o HA可能在乳腺癌淋巴管形态及增殖中发挥重要作用。

英文摘要：

This study aimed to investigate the change of hyaluronic acid in breast cancer and further to explore its effect on tight junction ZO-1 distribution and proliferation of lymphatic endothelial cells. Immunohistochemistry and enzyme linked immunosorbent assay were performed to test the expression of hyaluronic acid and serum concentration of hyaluronidase in benign breast disease and breast cancer, normal person and breast cancer, respectively. Immunofl uorescence, permeability of sodium fl uorescein, MTT and Western blot were applied to investigate the effects of HA and o HA on ZO-1 distribution, permeability of lymphatic endothelial cell monolayers, cell proliferation and the protein expression levels of ROCK1/Rho A. The results showed that the expressions of hyaluronic acid and hyaluronidase in breast cancer were higher than those of benign breast disease and normal person（P〈0.01）, respectively. Hyaluronic acid elevated with hyaluronidase in breast cancer, suggesting that the metabolism of HA was more activity. HA strengthened ZO-1 distribution on the cell membrane and unchanged the protein expression levels of ROCK1/Rho A. On the contrary, o HA induced ZO-1 distributed from cell membrane to cytoplasm, leading to disrupting the junction between cells, formed the intercellular space, stimulated permeability of cell monolayers, increased the protein expression levels of ROCK1/Rho A, and promoted proliferation of lymphatic endothelial cells（P〈0.01）. This study indicated that o HA might play an important role in lymphatic morphogenesis and lymphatic endothelial cell proliferation of breast cancer.