GRM5在人胶质瘤中表达及其与胶质瘤U251细胞迁移及侵袭能力关系的研究
发布时间：2019-01-11 13:10
【摘要】：研究目的:1.研究GRM5(代谢性谷氨酸受体5)在胶质瘤表达与胶质瘤病理级别的相关性;2.研究GRM5对胶质瘤U251细胞的迁移及侵袭能力的影响。研究方法:1.为了研究胶质瘤组织中GRM5表达情况,我们收集了本院神经外科13例胶质瘤标本,所有标本均依据2007年世界卫生组织(World Health Origanization,WHO)中枢神经系统肿瘤分类标准进行病理诊断并分级。应用免疫组化的方法检测胶质瘤标本中GRM5的表达情况。2.为了研究GRM5对胶质瘤U251细胞的迁移能力的影响,我们设计合成了2条特异性针对人GRM5基因的小分子干扰RNA(GRM5-siRNA-1247,GRM5-siRNA-1587),及对照RNA(GRM5-siRNA-NC);转染U251细胞株48h后用Western blot(蛋白质印迹法)检测GRM5的表达情况;3.用转染后的U251细胞做划痕实验和Tanswell实验,验证转染小分子RNA(GRM5-siRNA-1247,GRM5-siRNA-1587),干扰GRM5表达后U251细胞的迁移和侵袭能力的影响。实验结果:1.免疫组化结果显示GRM5在人低级别和高级别胶质瘤中均有表达,且随肿瘤级别增高,表达量也增高,结果具有统计学意义(P0.01)。2.Western Blot实验结果显示U251细胞转染小分子干扰RNA(GRM5-siRNA-1247)后GRM5表达量明显降低,差异具有统计学意义(t=3.020,P0.05)。3.划痕实验结果显示U251细胞转染小分子干扰RNA(GRM5-siRNA-1247)后,细胞划痕伤口愈合率明显低于对照组,差异具有统计学意义(24h t=6.751,P0.01;48h t=8.668 P0.01)。4.Tanswell实验结果显示U251细胞转染小分子干扰RNA(GRM5-siRNA-1247)后,细胞侵袭能力明显低于对照组,差异具有统计学意义(t=20.726,P0.01)。结论:1.GRM5在各级别胶质瘤中都有表达,且表达量与胶质瘤级别呈正相关。2.GRM5能促进胶质瘤U251细胞迁移及侵袭的能力。
[Abstract]:Objective: 1. To study the correlation between the expression of GRM5 (metabolic glutamate receptor 5) in gliomas and the pathological grade of gliomas. 2. To study the effect of GRM5 on the migration and invasion of U251 glioma cells. Methods: 1. In order to study the expression of GRM5 in glioma tissues, we collected 13 glioma specimens from neurosurgery department of our hospital, all of which were based on WHO (World Health Origanization, in 2007. WHO) Central nervous system tumor classification criteria for pathological diagnosis and grading. Immunohistochemical method was used to detect the expression of GRM5 in glioma specimens. 2. In order to study the effect of GRM5 on the migration ability of U251 glioma cells, we designed and synthesized two small interfering RNA (GRM5-siRNA-1247,GRM5-siRNA-1587) specific to human GRM5 gene and control RNA (GRM5-siRNA-NC). The expression of GRM5 was detected by Western blot (Western blot after transfection of U251 cell line for 48 h. The effect of small molecule RNA (GRM5-siRNA-1247,GRM5-siRNA-1587) transfection on the migration and invasion of U251 cells after GRM5 expression was tested by scratch test and Tanswell assay. Experimental results: 1. Immunohistochemical results showed that GRM5 was expressed in both low-grade and high-grade gliomas, and the expression level increased with the increase of tumor grade. The results showed that the expression of GRM5 in U251 cells was significantly decreased after transfection of small molecule interfering RNA (GRM5-siRNA-1247) (P0.01), and the difference was statistically significant (p0.05). The results of scratch test showed that after transfection of small molecule interfering RNA (GRM5-siRNA-1247), the wound healing rate of U251 cells was significantly lower than that of the control group (24 hours after transfection with small molecule interference RNA (P0.01). The results of 4.Tanswell assay showed that the invasion ability of U251 cells transfected with small molecule interfering RNA (GRM5-siRNA-1247) was significantly lower than that of the control group (tr 20.726 P0.01). Conclusion: the expression of 1.GRM5 is positively correlated with the grade of glioma, and 2.GRM5 can promote the migration and invasion of U251 cells.
【学位授予单位】：广州医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.41
本文编号：2407173
[Abstract]:Objective: 1. To study the correlation between the expression of GRM5 (metabolic glutamate receptor 5) in gliomas and the pathological grade of gliomas. 2. To study the effect of GRM5 on the migration and invasion of U251 glioma cells. Methods: 1. In order to study the expression of GRM5 in glioma tissues, we collected 13 glioma specimens from neurosurgery department of our hospital, all of which were based on WHO (World Health Origanization, in 2007. WHO) Central nervous system tumor classification criteria for pathological diagnosis and grading. Immunohistochemical method was used to detect the expression of GRM5 in glioma specimens. 2. In order to study the effect of GRM5 on the migration ability of U251 glioma cells, we designed and synthesized two small interfering RNA (GRM5-siRNA-1247,GRM5-siRNA-1587) specific to human GRM5 gene and control RNA (GRM5-siRNA-NC). The expression of GRM5 was detected by Western blot (Western blot after transfection of U251 cell line for 48 h. The effect of small molecule RNA (GRM5-siRNA-1247,GRM5-siRNA-1587) transfection on the migration and invasion of U251 cells after GRM5 expression was tested by scratch test and Tanswell assay. Experimental results: 1. Immunohistochemical results showed that GRM5 was expressed in both low-grade and high-grade gliomas, and the expression level increased with the increase of tumor grade. The results showed that the expression of GRM5 in U251 cells was significantly decreased after transfection of small molecule interfering RNA (GRM5-siRNA-1247) (P0.01), and the difference was statistically significant (p0.05). The results of scratch test showed that after transfection of small molecule interfering RNA (GRM5-siRNA-1247), the wound healing rate of U251 cells was significantly lower than that of the control group (24 hours after transfection with small molecule interference RNA (P0.01). The results of 4.Tanswell assay showed that the invasion ability of U251 cells transfected with small molecule interfering RNA (GRM5-siRNA-1247) was significantly lower than that of the control group (tr 20.726 P0.01). Conclusion: the expression of 1.GRM5 is positively correlated with the grade of glioma, and 2.GRM5 can promote the migration and invasion of U251 cells.
【学位授予单位】：广州医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.41
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