山姜素对U87人脑胶质瘤细胞的促凋亡作用及其机制研究

发布时间：2018-03-21 22:39

本文选题：胶质瘤　切入点：山姜素　出处：《青岛大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:胶质瘤是人最常见的颅内恶性肿瘤,它的发病率在全部的人脑肿瘤中占50%以上。而中低度分化的星形细胞瘤和胶质母细胞瘤的患者1年存活率不到全部患者的十分之一。人脑胶质瘤的快速增殖和低分化程度,较其他肿瘤相比较侵袭性强,仅只靠手术治疗仍无法做到完全切除,而手术切除后综合治疗仍是目前推荐的治疗方案。山姜素是一种天然的化合物,它可以抑制胶质瘤细胞的增殖和侵袭,并能诱导胶质瘤细胞发生细胞凋亡作用。本文主要研究山姜素对人脑胶质瘤细胞U87中bax,Bcl-2与Caspase,PARP蛋白的表达差异性和其促进凋亡作用的分子机制。山姜素则有望为人脑胶质瘤的治疗带来新的希望。方法:体外培养人脑胶质瘤细胞U87;通过MTT增殖实验法来检测山姜素对于U87细胞的增殖抑制作用;提取山姜素处理前后U87细胞总蛋白,Western blotting检测bax,Bcl-2与Caspase3,PARP的表达情况;通过流式细胞术来分析经过山姜素处理前后U87细胞的细胞周期变化;细胞划痕实验表明经过山姜素处理后的U87细胞迁移能力被明显抑制。结果:山姜素对U87细胞体外增殖具有显著的抑制作用并且这种抑制作用呈剂量依赖性,酶标仪测定在490 nm处的光密度即OD值,OD值可以反映出活细胞的数量。山姜素对胶质瘤U87细胞的增殖有显著的抑制作用,而且随着浓度的不断升高,U87细胞的存活率会逐渐下降,呈现出典型的浓度依赖性关系,其IC50为7.122μg/ml。;Westernblotting检测发现IC50剂量的山姜素处理U87细胞后,Caspase,bax的表达量明显高于对照组,而Bcl-2,PARP蛋白的表达量则明显的低于对照组,这提示山姜素可促进U87细胞凋亡;流式细胞术分析U87细胞在山姜素处理后细胞周期均呈现明显的差异,并且药物组细胞表现为G1期阻滞;Annexin V/PI实验显示经过山姜素处理后的U87细胞凋亡增加,并呈浓度依赖性。结论:山姜素可以抑制体外培养的人脑胶质瘤U87细胞的增殖作用,诱导其细胞凋亡相关基因的差异性表达,从而具有显著的促进细胞凋亡的作用,并能抑制U87细胞的迁移。然而,目前对细胞凋亡的认识认为,细胞的凋亡作用是一个非常复杂的,由多个基因调控的过程,山姜素在诱导人胶质瘤U87细胞凋亡的过程中是否有其它基因参与,以什么方式参与,仍有待于进一步的深入研究。
[Abstract]:Objective: glioma is the most common intracranial malignant tumor in human. Its incidence is more than 50% of all human brain tumors. The 1-year survival rate of patients with moderately and poorly differentiated astrocytoma and glioblastoma is less than 1/10 of all patients. More aggressive than other tumors, it is still impossible to completely remove the tumor by surgical treatment alone, and comprehensive treatment after surgical resection is still the recommended treatment. Curcumin is a natural compound. It can inhibit the proliferation and invasion of glioma cells. The effect of curcumin on the expression of baxbu2 and caspase- PARP in human glioma cell U87 and the molecular mechanism of its role in promoting apoptosis were studied in this paper. Methods: human glioma cell U87 was cultured in vitro, and the inhibitory effect of curcumin on U87 cell proliferation was detected by MTT proliferation assay. The expression of bcl-2 and Caspase3PARP in U87 cells was detected by Western blotting before and after treatment with curcumin, and the cell cycle changes of U87 cells were analyzed by flow cytometry. Cell scratch assay showed that the migration ability of U87 cells treated with curcumin was significantly inhibited. Results: the proliferation of U87 cells was significantly inhibited by curcumin in a dose-dependent manner. The optical density at 490 nm, OD value and OD value, can reflect the number of living cells. Curcumin can inhibit the proliferation of U87 cells, and the survival rate of U87 cells decreases with increasing concentration. There was a typical concentration-dependent relationship between the two groups. The IC50 was 7.122 渭 g / ml. Western blotting analysis showed that the expression of CaspaseB Bax in U87 cells treated with IC50 was significantly higher than that in the control group, while the expression of Bcl-2PARP protein was significantly lower than that in the control group. These results suggested that curcumin could promote the apoptosis of U87 cells, and that the cell cycle of U87 cells treated with curcumin was significantly different by flow cytometry. In the drug group, the G1 phase arrest Annexin V/PI assay showed that the apoptosis of U87 cells treated with curcumin increased in a concentration dependent manner. Conclusion: curcumin can inhibit the proliferation of human glioma U87 cells cultured in vitro. The differential expression of apoptosis-related genes induced by U87 cells can significantly promote apoptosis and inhibit the migration of U87 cells. Apoptosis is a complex process regulated by multiple genes. Whether and how other genes are involved in the process of inducing apoptosis of human glioma U87 cells need to be further studied.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.41

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