hIGF-1诱导和aggrecanse-1沉默对人骨髓干细胞微丝结构的影响

发布时间：2018-01-30 09:38

本文关键词： hIGF-1 aggrecanse-1 转染骨髓干细胞微丝　出处：《青岛大学》2016年硕士论文　论文类型：学位论文

【摘要】：目的探讨h IGF-1(人胰岛素生长样因子-1)诱导和aggrecanse-1(多聚蛋白聚糖酶-1)沉默对人骨髓干细胞微丝结构的影响。方法用基因重组技术构建携带h IGF-I过表达和沉默aggrecanase-1-si RNA的慢病毒载体,用其转染人骨髓干细胞,记为基因诱导组、基因沉默组、基因诱导沉默组,以不携带目的基因的慢病毒转染人骨髓干细胞和未处理的人骨髓干细胞分别记为对照组1、对照组2,转染7天后PCR检测基因表达,同时激光共聚焦显微镜下观察各组细胞的微丝结构,并用Image J检测各组细胞的荧光强度。用SPSS软件进行统计学分析。结果基因诱导沉默组的荧光强度(0.119±0.008)高于基因诱导组(0.081±0.001)和基因沉默组(0.080±0.003),基因诱导组和基因沉默组的荧光强度高于对照组1(0.065±0.003)和对照组2(0.066±0.005),差异有统计学意义(P0.01)。对照组1和对照组2的荧光强度差异无统计学意义(P0.05)。结论h IGF-1的诱导和aggrecanse-1的沉默均促进骨髓干细胞微丝结构的生成,诱导沉默双向作用更显著。
[Abstract]:Objective to investigate the induction of hIGF-1 (human insulin-like factor-1) and aggrecanse-1 (PC-1). Methods Lentivirus vector carrying h IGF-I overexpression and silencing aggrecanase-1-si RNA was constructed by gene recombination technique. Human bone marrow stem cells were transfected with them as gene induction group, gene silencing group and gene induced silencing group. Human bone marrow stem cells and untreated human bone marrow stem cells transfected with lentivirus without the target gene were recorded as control group 1 and control group 2 respectively. After 7 days of transfection, gene expression was detected by PCR. At the same time, the microfilament structure of each group was observed under confocal laser microscope. The fluorescence intensity of each group was detected by Image J and analyzed statistically by SPSS software. Results the fluorescence intensity of gene induced silencing group was 0.119 卤0.008). It was higher than that of gene induction group (0.081 卤0.001) and gene silencing group (0.080 卤0.003). The fluorescence intensity of gene induced group and gene silencing group was higher than that of control group (1 0. 065 卤0. 003) and control group (2. 066 卤0. 005). There was no significant difference in fluorescence intensity between control group 1 and control group 2 (P0.05). Conclusion both h IGF-1 induction and aggrecanse-1 silencing can promote the formation of bone marrow stem cell microfilaments. The bidirectional effect of induced silencing is more significant.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R684

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