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目的探讨白细胞介素8(IL-8)在乳腺癌细胞中促血管生成作用,及其与雌激素受体之间的关系。方法收集不同的乳腺癌细胞培养上清液,通过观察这些上清液对人脐静脉内皮细胞迁移、增殖和新生毛细血管管腔形成三个体外血管生成实验以及裸鼠体内血管生成实验来研究细胞因子在乳腺癌血管生成中的作用机制;采用细胞上清液芯片实验检测MDA-MB-231细胞和雌激素受体稳定转染的MDA-MB-231细胞的IL-8表达水平;进一步将雌激素受体和IL-8启动子瞬时转染至MDA-MB-231细胞,经双荧光素酶报告基因检测雌激素受体对IL-8的调控作用。结果与IL-8低表达细胞相比较,IL-8高表达和中表达细胞上清液所培养的HUVEC更易于发生迁移(t值分别为6.94和17.75,P均<0.05),IL-8抗体可以特异性地阻断这种作用(t值分别为16.67和9.08,P均<0.05)。雌激素受体阴性MDA-MB-231细胞经过雌激素受体稳定转染后,IL-8的表达水平下降8.8倍。雌激素受体α能够下调IL-8启动子的活性(r=0.856,P<0.05),对照载体无此作用。结论IL-8具有促乳腺癌细胞血管生成作用,乳腺癌细胞中IL-8的水平与雌激素受体水平呈负相关,外源性的雌激素受体可下调乳腺癌细胞中的IL-8表达。
Objective To investigate the effect of interleukin-8 (IL-8) on angiogenesis in breast cancer cells and its relationship with estrogen receptor. Methods Different breast cancer cell culture supernatants were collected and the effects of these supernatants on migration, proliferation and neovascularization of human umbilical vein endothelial cells were observed in vitro and in vivo. Factor in mammary gland angiogenesis. IL-8 expression in MDA-MB-231 cells stably transfected with MDA-MB-231 cells and estrogen receptor was detected by cell supernatant chip assay. The hormone receptor and IL-8 promoter were transiently transfected into MDA-MB-231 cells. The dual luciferase reporter gene was used to detect the regulatory effect of estrogen receptor on IL-8. Results Compared with IL-8 low expression cells, HUVECs cultured in supernatant of IL-8 and medium-expressed cells were more likely to migrate (t values were 6.94 and 17.75, P <0.05 respectively), while IL-8 antibody This effect can be specifically blocked (t values were 16.67 and 9.08, P <0.05 respectively). After stable transfection of estrogen receptor in estrogen receptor-negative MDA-MB-231 cells, the expression level of IL-8 decreased 8.8-fold. Estrogen receptor alpha decreased the activity of the IL-8 promoter (r = 0.856, P <0.05), whereas the control vector did not. Conclusion IL-8 can promote the angiogenesis of breast cancer cells. The level of IL-8 in breast cancer cells is negatively correlated with the level of estrogen receptor. Exogenous estrogen receptor can down-regulate the expression of IL-8 in breast cancer cells expression.