褐藻糖胶对U266细胞自噬、迁移和侵袭的影响

发布时间：2018-03-24 19:13

本文选题：褐藻糖胶　切入点：多发性骨髓瘤　出处：《重庆医科大学》2017年硕士论文

【摘要】：目的多发性骨髓瘤(multiple myeloma,MM)的发病率在血液肿瘤中排名第三。尽管化疗方案的改善、新药的应用及干细胞移植技术的成熟使该疾病治疗缓解率大大提高,但相当一部分患者在数年或数月内复发。有研究表明微小残留病灶(minimal residual disease,MRD)引起的复发是其难以治愈的原因之一。因此,发现能够抑制骨髓瘤细胞迁移、侵袭能力,减少MRD的低毒性药物,对辅助治疗有着重要的作用。本研究探讨褐藻糖胶对多发性骨髓瘤U266细胞自噬、迁移及侵袭的影响,为研究褐藻糖胶治疗多发性骨髓瘤的临床应用提供理论依据。方法褐藻糖胶处理细胞后,透射电子显微镜(TEM)观察细胞自噬小体,Traswell小室观察细胞迁徙及侵袭能力,Western Blot测定LC3-Ⅱ/LC3-Ⅰ,Beclin-1,P62,MMP9,CXCR4,p-AKT/T-AKT,p-mTOR/T-mTOR蛋白表达,ELISA法检测细胞培养液中MMP9含量。结果(1)TEM观察发现,褐藻糖胶处理组自噬小体增加。(2)褐藻糖胶可降低U266细胞迁移及侵袭能力,并呈现出剂量依赖性,氯喹可阻断该抑制作用。(3)Western Blot结果显示,LC3-II/LC3-I、Beclin-1及MMP9蛋白表达随褐藻糖胶浓度增加而升高,P62、CXCR4、p-AKT/T-AKT及p-mTOR/T-mTOR蛋白表达与褐藻糖胶浓度增加而降低。(4)ELISA结果显示,褐藻糖胶处理组细胞培养液中MMP9的含量下降。结论褐藻糖胶可诱导多发性骨髓瘤U266细胞自噬,抑制该细胞的迁移、侵袭能力。
[Abstract]:Objective multiple myeloma (MM) is the third most common disease in blood tumors. Despite the improvement of chemotherapy regimen, the application of new drugs and the maturity of stem cell transplantation technology, the remission rate of the disease has been greatly improved. However, a considerable number of patients recurred within a few years or months. Some studies have shown that the recurrence caused by minimal residual disease is one of the reasons for the difficulty of cure. Therefore, it has been found that it can inhibit the migration and invasion ability of myeloma cells. Reducing the low toxicity of MRD plays an important role in adjuvant therapy. In this study, we investigated the effects of fucoidan on autophagy, migration and invasion of multiple myeloma U266 cells. Methods to study the clinical application of fucoidan in the treatment of multiple myeloma. The transmissive electron microscopy (TEM) was used to observe the migration and invasiveness of the cells in the autophagy chamber. Western Blot was used to detect the expression of CXCR4, p-AKTR, p-mTOR-T-mTOR protein in the cell culture medium. In the treatment group, the autophagy increased. 2) Alginate decreased the migration and invasion ability of U266 cells in a dose-dependent manner. Chloroquine blocked the inhibitory effect. Western Blot results showed that the protein expression of Beclin-1 and MMP9 increased with the increase of the concentration of fucoidin, and the expression of P62CXCR4 p-AKT and p-mTOR/T-mTOR protein decreased with the increase of the concentration of fucoidan. Conclusion Alginate can induce autophagy of multiple myeloma U266 cells and inhibit the migration and invasion of U266 cells.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R733.3

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