慢性缺氧应激可能通过EMT增强乳腺癌MCF-7细胞恶性生物学行为

发布时间：2018-05-07 12:15

本文选题：乳腺癌 + 缺氧　；参考：《中国病理生理杂志》2017年10期

【摘要】：目的:探讨慢性缺氧应激对人乳腺癌MCF-7细胞恶性生物学行为的影响及可能机制。方法:将人乳腺癌MCF-7细胞分为缺氧组(1%O_2、5%CO_2和94%N_2)和正常对照组(常氧)进行培养。利用MTT法、CCK-8实验、细胞直接计数法及细胞侵袭和迁移实验对MCF-7细胞活力、增殖及侵袭和迁移能力进行检测;用软琼脂集落形成实验及Matrigel 3D培养技术检测MCF-7细胞非锚定生长能力及极性改变情况;利用MCF-7细胞构建裸鼠皮下种植瘤模型,检测慢性缺氧应激对体内肿瘤生长及肺转移的影响;利用倒置显微镜观察MCF-7细胞形态改变;Western blot检测低氧诱导因子1(hypoxia-inducible factor-1,HIF-1)和磷酸化的糖原合成酶激酶3β(glycogen synthase kinase-3β,GSK-3β)在缺氧环境下表达水平的改变,以及E-钙黏连蛋白(E-cadherin)、N-钙黏连蛋白(Ncadherin)、波形蛋白(vimentin)、基质金属蛋白酶3(matrix metalloproteinase-3,MMP-3)、MMP-9等上皮-间充质转化(epithelial-mesenchymal transition,EMT)相关蛋白的表达水平。结果:与正常对照组相比较,慢性缺氧组MCF-7细胞活力、增殖能力及侵袭迁移能力增强,细胞非锚定生长能力提高且在3D培养系统更容易发生极性改变,呈现侵袭样生长,体内生长及转移能力增强;除了HIF-1被缺氧诱导表达升高外,GSK-3β呈现活化趋势,且上皮样标志物E-cadherin蛋白表达水平明显下降,而间充质样标志物N-cadherin、vimentin、MMP-3和MMP-9蛋白表达水平明显升高。结论:慢性缺氧应激促进了乳腺癌细胞恶性生物学行为,且其机制可能与EMT有关。
[Abstract]:Objective: to investigate the effect of chronic hypoxia stress on malignant biological behavior of human breast cancer MCF-7 cells and its possible mechanism. Methods: human breast cancer MCF-7 cells were divided into hypoxia group and normal control group. The viability, proliferation, invasion and migration of MCF-7 cells were detected by MTT assay, direct cell count and cell invasion and migration assay. Soft Agar colony forming test and Matrigel 3D culture were used to detect the unanchored growth ability and polarity change of MCF-7 cells, the subcutaneous tumor model of nude mice was constructed by MCF-7 cells, and the effects of chronic hypoxia stress on tumor growth and lung metastasis in vivo were examined. The morphological changes of MCF-7 cells were observed by inverted microscope. The expression of hypoxia inducible factor 1(hypoxia-inducible factor-1 (HIF-1) and phosphorylated glycogen synthase kinase 3 尾 -glycogen synthase kinase-3 尾 -GSK-3 尾 were detected by Western blot under hypoxia. The expression levels of E-cadherin, vimentin, 3(matrix metalloproteinase-3 and MMP-3MMP 9 in epithelial-mesenchymal transition (EMT9) were also found in Ecadherin (E-cadherin), vimentin (vimentin), matrix metalloproteinase-3 (MMP) and matrix metalloproteinase-3 (MMPs). Results: compared with the control group, the MCF-7 cell viability, proliferation and invasion and migration ability of chronic hypoxia group were increased, and the cell unanchored growth ability was increased. In 3D culture system, polarity changes were more likely to occur, showing invasive growth. The ability of growth and metastasis in vivo was enhanced, the expression of GSK-3 尾 was increased by hypoxia, and the expression level of E-cadherin protein decreased significantly, while the expression of N-cadherin protein MMP-3 and MMP-9 protein of mesenchymal marker N-cadherin protein increased significantly. Conclusion: chronic hypoxia stress promotes malignant biological behavior of breast cancer cells, and its mechanism may be related to EMT.
【作者单位】：安徽中医药大学;厦门大学第一附属医院超声科;安徽医科大学;
【基金】：中国博士后科学基金第59批面上资助项目(No.2016M592037) 安徽省自然科学基金资助项目(No.1608085MH187)
【分类号】：R737.9

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