Syndecan-1对syntenin表达及胶质瘤细胞迁移能力的影响

发布时间：2018-03-01 16:38

本文关键词： syndecan-1 syntenin 胶质瘤迁移 Rac1　出处：《遵义医学院》2017年硕士论文　论文类型：学位论文

【摘要】：目的:探讨syndecan-1和syntenin在胶质瘤中的表达及其相互作用对胶质瘤细胞迁移能力的影响。方法:利用免疫组化分析syndecan-1和syntenin在胶质瘤中的表达部位、表达水平,与胶质瘤级别的关系及二者在胶质瘤中的表达相关性;构建syndecan-1过表达质粒及小干扰RNA序列分别转染胶质瘤u251细胞,western blot检测syndecan-1、syntenin、总的Rac1的表达;细胞划痕实验和Transwell实验观察不同处理组细胞的迁移能力;Rac1活性实验检测不同处理组活化的Rac1蛋白水平。结果:高恶性级别胶质瘤中,syndecan-1和syntenin表达水平较低恶性级别及对照组明显增强(H=12.784,P=0.002;H=16.274,P=0.000),syndecan-1与syntenin表达均与胶质瘤恶性级别呈正相关(rs=0.576,P=0.001;rs=0.661,P=0.000),且syndecan-1与syntenin在胶质瘤组织中的表达存在正相关(rs=0.628,P=0.001)。SDC1过表达的u251细胞中,syndecan-1、syntenin蛋白水平明显高于对照及空载组(F=83.216,P=0.000;F=15.571,P=0.004),SDC1沉默组中,syndecan-1、syntenin蛋白水平明显低于对照及阴性组(F=60.728,P=0.000;F=16.747,P=0.004);Transwell实验发现:与对照组相比,SDC1过表达可明显增强胶质瘤细胞迁移能力(F=95.559,P=0.000),而沉默SDC1后胶质瘤细胞迁移能力明显减弱(F=111.391,P=0.000);细胞划痕实验显示:与对照组相比,SDC1过表达可明显增强胶质瘤细胞划痕修复能力(F=69.89,P=0.000),而沉默SDC1后胶质瘤细胞划痕修复能力减弱(F=50.819,P=0.000);Rac1活性实验发现:SDC1过表达后胶质瘤细胞Rac1活化水平明显高于对照及空载组(F=12.37,P=0.007),沉默SDC1后Rac1活化水平明显低于对照及阴性组(F=13.082,P=0.006),各处理组总的Rac1无明显改变(P0.05)。结论:1、syndecan-1和syntenin在正常脑组织中表达较少或缺如,在胶质瘤组织中随胶质瘤级别增高而表达增高,且两基因在胶质瘤中存在正相关;2、syndecan-1过表达上调syntenin的表达,进一步通过激活Rac1促进胶质瘤细胞迁移。
[Abstract]:Objective: To investigate the effect of expression of syndecan-1 and syntenin in glioma and their interaction on the migration of glioma cells. Methods: using immunohistochemical analysis of the expression site, syndecan-1 and syntenin in glioma expression, expression in gliomas and the relationship between the level of glioma and two syndecan-1 construction; over expression of plasmid RNA and small interfering sequences were transfected into U251 glioma cells, Western blot detection of syndecan-1, syntenin, total Rac1; migration of cell scratch experiments and Transwell experiments of cells; Rac1 protein level Rac1 activity assay in different groups. Results: the activation of high grade gliomas the expression of syntenin, syndecan-1 and the low level of malignant level and control group increased significantly (H=12.784, P=0.002; H=16.274, P=0.000), syndecan-1 and syntenin expression with glue Related to tumor malignant level positively (rs=0.576, P=0.001; rs=0.661, P=0.000), and there was a positive correlation between syndecan-1 and syntenin expression in glioma tissues (rs=0.628, P=0.001) syndecan-1.SDC1 overexpression in U251 cells, the protein level of syntenin was significantly higher than the control group and no load (F=83.216, P=0.000; F=15.571, P=0.004). Syndecan-1 SDC1 silencing group, syntenin protein levels were significantly lower than the control, and the negative group (F=60.728, P=0.000; F=16.747, P=0.004; Transwell) found that: compared with the control group, SDC1 expression significantly increased glioma cell migration (F=95.559, P=0.000), and the silencing of SDC1 glioma cell migration ability decreased significantly (F=111.391, P=0.000); cell scratch test: compared with the control group, SDC1 overexpression can significantly enhance glioma cell scratch repair ability (F=69.89, P=0.000), and the silencing of SDC1 glioma cell scratch repair The complex capacity decreased (F=50.819, P=0.000); Rac1 activity test: overexpression of SDC1 in glioma cells Rac1 activation level is significantly higher than the control group and no load (F=12.37, P=0.007), SDC1 silencing Rac1 activation levels were significantly lower than the control group and the negative group (F=13.082, P=0.006), each treatment group had no significant change (Rac1 P0.05). Conclusion: 1. The expression of syndecan-1 and syntenin in small or absent in normal brain tissue in glioma with glioma grade increased expression and two genes in gliomas has a positive correlation; 2, overexpression of syndecan-1 up-regulated the expression of syntenin, further through the activation of Rac1 promote glioma cell migration.

【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.41

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