调控内源性大麻素系统对不同脑胶质瘤细胞的影响研究

发布时间：2018-02-11 04:50

本文关键词： 内源性大麻素系统胶质瘤细胞雷公藤红素　出处：《厦门大学》2014年硕士论文　论文类型：学位论文

【摘要】：研究背景：脑胶质瘤是恶性度很高、易复发、预后差的颅内恶性肿瘤之一,在临床上一直是个难题。许多研究证明内源性大麻素具有抗肿瘤生成作用,包括抑制增殖、诱导凋亡和抑制迁移作用,但其对胶质瘤的作用靶点及详细机制尚未完全阐明。雷公藤红素可以抑制蛋白酶体活性、阻滞在细胞周期和诱导细胞凋亡,这使得它成为一种潜在的治疗胶质瘤的药物。目的：本研究通过观察利用单独分子生物学、分子药物学手段调控内源性大麻素系统或与雷公藤红素联用后不同胶质瘤细胞的增殖情况,在细胞水平探讨胶质瘤可能的治疗靶点,为雷公藤红素开发成为抗胶质瘤新药提供依据。方法：用细胞计数板计算细胞数绘制细胞的生长曲线,根据曲线计算出倍增时间；用Western blot方法检测细胞内CD133表达；内源性大麻素、游离脂肪酸及神经酰胺采用LC-MS/MS方法检测;使用荧光定量PCR (RT-qPCR)技术检测大麻素受体CB1、CB2,大麻素水解酶FAAH、MGL等的基因mRNA表达水平及雷公藤红素对其的影响；用CCK-8试剂盒检测单独调控内源性大麻素系统或与雷公藤红素联用后胶质瘤细胞增殖的影响。结果：SHG细胞增长速度最快,具有干细胞特征。四种细胞内FFAs、Ceramides含量均有显著性差异。内源性大麻素系统中大麻素AEA、2-AG及其水解酶FAAH、MGL,以及CB1的表达有显著性差异,2-AG和MGL是调控内源性大麻素系统的重要靶点。雷公藤红素对胶质瘤细胞具有一定程度的抑制作用,存在时间和剂量依赖性,在此过程中上调了大麻素受体的基因表达,CB2的升高更为明显,雷公藤红素与2-AG分解酶抑制剂JZL184、小剂量的大麻素受体激动剂都可以增强的抗胶质瘤作用,可以作为抗胶质瘤作用的联合用药开发。结论：上调内源性大麻素2-AG含量或激活相关信号通道会抑制胶质瘤细胞的增殖,内源性大麻素系统是治疗胶质瘤的有效靶点,雷公藤红素作为抗胶质瘤联合用药开发具有良好的临床应用前景。
[Abstract]:Background: glioma is one of intracranial malignant tumors with high degree of malignancy, easy recurrence and poor prognosis, which has been a difficult problem in clinic. Many studies have proved that endogenous cannabinoids have anti-tumorigenic effects, including inhibition of proliferation. However, the target and detailed mechanism of its action on glioma has not been fully elucidated. Tripterygium wilfordii can inhibit proteasome activity, block cell cycle and induce apoptosis. This makes it a potential therapy for glioma. Objective: to investigate the effects of monolayer molecular biology and molecular pharmacology on the proliferation of glioma cells with endogenous cannabinoid system or tripterygium wilfordii. To explore the possible therapeutic targets of glioma at cell level and to provide evidence for the development of tripterine as a new anti-glioma drug. Methods: the cell growth curve was calculated by cell counting board, and the doubling time was calculated according to the curve. The expression of CD133 was detected by Western blot method, and the endogenous cannabinoid, free fatty acid and ceramide were detected by LC-MS/MS method. The level of mRNA expression of cannabinoid receptor CB1 (CB1) and cannabinase (FAAHH) MGL and the effect of tripterine on it were detected by fluorescence quantitative PCR RT-qPCR technique. The effects of endogenous cannabinoid system alone or tripterygium wilfordii on the proliferation of glioma cells were detected by CCK-8 kit. The results showed that the growth rate of the cell was the fastest. There were significant differences in the contents of FFAs-Ceramides in four kinds of cells. The expression of cannabinoid AEA-2-AG and its hydrolase FAAHMAMGLand CB1 in endogenous cannabinoid system were significantly different, and 2-AG and MGL regulated the endogenous cannabinoid system. Tripterygium wilfordii has a certain degree of inhibitory effect on glioma cells. In a time-and dose-dependent manner, the gene expression of cannabinoid receptor (CB2) was upregulated in a dose-dependent manner. Tripterygium wilfordii and 2-AG decomposing enzyme inhibitor JZL1844, a small dose of cannabinoid receptor agonist, could enhance the anti-glioma effect. It can be used as a combined drug for anti-glioma. Conclusion: upregulation of endogenous cannabinoid 2-AG or activation of related signal channels can inhibit the proliferation of glioma cells. Endogenous cannabinoid system is an effective target for the treatment of glioma. Tripterygium wilfordii as a combined antiglioma drug has a good clinical application prospects.
【学位授予单位】：厦门大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.4

【共引文献】