中文摘要：

目的：观察shRNA介导的Y-盒结合蛋白-1（YB-1）表达沉默对神经母细胞瘤细胞体内成瘤能力的影响。方法：构建针对YB-1的小干扰RNA（siRNA）真核表达载体,利用LipofectamineTM2000脂质体转染人神经母细胞瘤SH-SY5Y细胞。Western blot技术检测干扰后YB-1蛋白表达水平变化。平板克隆形成实验检测YB-1蛋白受抑是否影响细胞凋亡。通过测量肿瘤体积和重量评估YB-1蛋白受抑对裸鼠体内成瘤的影响。HE染色和TUNEL实验评估干扰对肿瘤细胞增殖及凋亡的影响。结果：Western blot技术检测无效转染组（NC组）与对照组（SH-SY5Y组）相比,YB-1表达量无显著差异,而转染组（YB-1 shRNA组）与SH-SY5Y组相比,YB-1表达水平明显下降。克隆形成实验显示NC组细胞克隆形成能力与SH-SY5Y组细胞相比未发生显著变化（P〉0.05）,而与SH-SY5Y组相比,YB-1 shRNA组克隆形成能力显著下降（P〈0.01）。裸鼠成瘤统计分析结果显示,SH-SY5Y组的平均肿瘤体积和平均肿瘤重量明显高于YB-1 shRNA组（P〈0.01）,NC组肿瘤体积与重量与SH-SY5Y组相比无明显差异（P〉0.05）。HE染色和TUNEL染色发现SH-SY5Y组和NC组细胞与YB-1 shRNA组相比分裂活跃,凋亡细胞较少。结论：shRNA介导的YB-1表达沉默抑制细胞生长,诱导凋亡,显著抑制神经母细胞瘤的体内成瘤能力。

英文摘要：

Objective：To investigate the influence of shRNA- mediated silencing of Y- Box Binding Protein- 1（ YB- 1） on tumorigenesis of neuroblastoma cell SH- SY5 Y in vivo. Methods：Establishing the small interfering RNA（ siRNA） eukaryotic expression vector targeting YB- 1,transfection was performed using LipofectamineTM2000 liposomeinto human neuroblastoma SH- SY5 Y cells. The expression of YB- 1 protein was detected with Western blot after interference. The effect of YB- 1 protein after inhibition on cell apoptosis was detected by plate clone formation assay. The effect of YB- 1 protein after inhibition on tumor growth in nude mice was estimated by measuring the size of tumor volume and weight. The effect of interferenceon the proliferation and apoptosis of tumor cells was assessed by HE staining and TUNEL analysis. Results：Compared with invalid transfection group（ NC group）,the expression of YB- 1 of control group（ SH- SY5 Y group） had no significant difference by Western blot,however,the expression level of YB- 1 of the transfection group（ YB- 1 shRNA group） decreased significantly compared with the SH-SY5 Y group,clone formation assay showed that colony formation ability of NC group cells compared with SH- SY5 Y group cells did not change significantly（ P 0. 05）,but YB- 1 shRNA group colony formation ability decreased significantly compared to the SH- SY5 Y group（ P 0. 01）. Statistical analysis of tumor growth in nude mice showed that the average tumor volume and average tumor weight of SH- SY5 Y group was significantly higher than those of YB- 1 shRNA group（ P 0. 01）,there was no difference about tumor volume and weight between NC group and SH- SY5 Y group（ P 0. 05）. HE staining and TUNEL analysis revealed that SH- SY5 Y group and NC group compared with YB- 1 shRNA group had more active cell division ability and fewer apoptotic. Conclusion：shRNA- mediated silencing of YB- 1 led to a growth arrest and to induction of apoptosis,as well as significantly inhibited tumorigenesis