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RNAi NRP-1基因对骨肉瘤细胞凋亡、ROS含量及STAT3信号通路的影响
作者:韩耀光1  燕太强2  潘富文1  邓睿1 
单位:1. 海南省第三人民医院 骨科, 海南 三亚 572000;
2. 北京大学人民医院 骨肿瘤科, 北京 100044
关键词:NRP-1基因 骨肉瘤 凋亡 ROS含量 STAT3信号通路 
分类号:R738.6
出版年·卷·期(页码):2019·38·第一期(1-5)
摘要:

目的:探讨RNAi神经纤毛蛋白1(NRP1)基因对骨肉瘤细胞凋亡、ROS含量及STAT3信号通路的影响。方法:分别将合成的NRP-1的siRNA(NRP-1-siRNA组)及无干扰作用的siRNA(NC组)转染人骨肉瘤MG-63细胞,并设置空白对照组,Western blotting检测转染48 h的细胞中NRP-1和磷酸化的STAT3(p-STAT3)及STAT3信号通路靶基因Cyclin D1和Bcl-2的蛋白表达;CCK8法分别于转染的24、48 h和72 h检测细胞活力;流式细胞术分别检测细胞凋亡率及ROS含量。结果:与空白对照组比较,NRP-1-siRNA组NRP-1、Cyclin D1、Bcl-2和p-STAT3的蛋白表达均显著降低,转染24、48、72 h的细胞活力均显著降低,细胞凋亡率显著升高,ROS含量显著升高(P<0.05)。结论:RNAi抑制NRP-1基因表达可降低骨肉瘤细胞活力,诱导细胞凋亡,机制可能与提高细胞内ROS含量和下调STAT3信号通路有关。

Objective:To investigate the effect of NRP-1 gene by RNA interference on apoptosis, ROS content and STAT3 signaling pathway in osteosarcoma cells. Methods:siRNA of synthetic NRP-1 (NRP-1-siRNA group) and siRNA without interference (NC group) were transfected to human osteosarcoma MG-63 cells, respectively.Western blotting was used to detect the expression of NRP-1, STAT3, phosphorylated STAT3(p-STAT3) and STAT3 signaling pathway target genes Cyclin D1 and Bcl-2 protein cells transfected for 48 h;CCK8 assay was used to detect cell viability cells transfected 24 h, 48 h and 72 h, respectively; cell apoptosis rate and ROS content were detected by flow cytometry. Results:Compared with the blank control group, the protein expression of NRP-1, Cyclin D1, Bcl-2 and p-STAT3 in NRP-1-siRNA group decreased significantly, and the cell viability decreased significantly at three time points, the apoptosis rate and ROS content increased significantly (P<0.05). Conclusion:Inhibition of NRP-1 gene expression by RNA interference can reduce osteosarcoma cell viability, induce cell apoptosis, the mechanism may be related to increase ROS content and downregulate STAT3 signaling pathway.

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