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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。
The purpose of this study was to investigate the effect of hyaluronic acid (HA) molecular size change on the distribution of ZO-1 (zonula occludens-1) and cell proliferation in breast cancer. Immunohistochemistry was used to detect the expression of HA in 8 cases of benign breast disease and breast cancer tissues. The serum hyaluronidase (Hyase) levels in 20 pairs of normal and breast cancer patients were detected by enzyme-linked immunosorbent assay (ELISA) The effects of HA and oligosaccharides of Hyaluronic acid (o HA) on the expression of ZO-1, monolayer permeability were observed by immunofluorescence, sodium fluorescein permeation assay, MTT proliferation assay and Western blot respectively. , Cell proliferation and downstream ROCK1 / Rho A signaling pathways. The results showed that compared with benign breast disease control group, the expression of HA in breast cancer patients was significantly increased (P <0.01). Compared with normal subjects, serum Hyase levels were significantly increased in breast cancer patients (P <0.01), suggesting breast cancer When, HA increased with its degradation enzyme increased, HA metabolic activity increased. HA enhanced the connection between ZO-1 cells and had no significant effect on the expression of ROCK1 / Rho A protein; on the contrary, o HA promoted the distribution of ZO-1 in lymphatic endothelial cells from cytoplasm to cytoplasm, Clearance and lymphatic vessel permeability, upregulated the expression of ROCK1 / Rho A protein and promoted the proliferation of lymphatic endothelial cells (P <0.01). Research suggests that o HA may play an important role in lymphatic vessel morphology and proliferation in breast cancer.