乳腺癌相关透明质酸变化对淋巴内皮细胞紧密连接分子ZO-1分布及.PDF

中国细胞生物学学报 Chinese Journal of Cell Biology 2015, 37(2): DOI: 10.11844/cj cb.2015.02.0266 乳腺癌相关透明质酸变化对淋巴内皮细胞紧密连接 分子ZO-1分布及细胞增殖的影响 余梦思 刘 雯 何怡青 杨翠霞 杜 艳 吴 曼 张国良 高 锋* (上海交通大学附属第六人民医院中心实验室, 上海200233) 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, oHA)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ROCK 1/RhoA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(P0.01); 与 正常人相比, 乳腺癌患者血清Hyase水平显著升高(P0.01), 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ROCK 1/RhoA蛋白表达水平无明显影响; 相反, oHA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ROCK 1/RhoA蛋白表达水平, 促进淋巴内皮细胞增殖(P0.01) 。该研究提示, oHA可能在乳腺癌淋巴管形态及增殖中发挥重要作用。 关键词 乳腺癌; oHA; 淋巴内皮细胞; ZO-1; 增殖 The Variation of Hyaluronic Acid in Breast Cancer and Its Effect on Tight Junction ZO-1 and Proliferation of Lymphatic Endothelial Cells Yu Mengsi, Liu Yiwen, He Yiqing, Yang Cuixia, Du Yan, Wu Man, Zhang Guoliang, Gao Feng* (Department of Central Laboratory, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital, Shanghai 200233, China) Abstract 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. Immunofluorescence, permeability of sodium fluorescein, MTT, Western blot were applied to inv