癌变·畸变·突变2017，Vol.29Issue(5)：329-334,339,7.DOI:10.3969/j.issn.1004-616x.2017.05.002
慢病毒介导的TPX2基因沉默对人宫颈癌HeLa细胞系增殖、迁移和细胞周期的影响及机制
Effect and mechanism of lentivirus-mediated silencing of TPX2 gene expression on proliferation, migration and cell cycle of HeLa cells
摘要
Abstract
OBJECTIVE: To investigate the effect and mechanism of silencing of TPX2 gene expression by lentivirus on cell proliferation,migration and cell cycle in human cervical cancer HeLa cells. METHODS:Four lentivirus expression vectors carrying TPX2-shRNA1/2/3/4 which target TPX2 gene (named as LV-TPX2-shRNA-1/2/3/4) were constructed,while vector TPX2-NC-shRNA was used as the negative control. The five plasmids were separately transfected into 293T cells to prepare the recombinant lentivirus. The recombinant lentivirus carrying TPX2-shRNA1/2/3/4 and TPX2-NC-shRNA were infected into HeLa cells,respectively. Then,expression levels of TPX2 mRNA and protein in HeLa cells were determined by real-time fluorescent quantitative PCR and Western blotting,respectively. The effect of TPX2 gene silencing on the proliferation of HeLa cells was detected using the CCK-8 method. Cell migration and cell-cycle distribution were detected by transwell migration experiment and flow cytometry,respectively,in each group of HeLa cells. Changes of Ki-67, cyclin B2, P53 and Aurora-A expressions were detected by Western blotting. RESULTS:Compared with the negative control group,expressions of TPX2 mRNA and protein in HeLa cells were significantly inhibited by LV-TPX2-shRNA, especially LV-TPX2-shRNA-1 (P<0.01). Subsequently, LV-TPX2-shRNA-1 was selected for follow-up experiments. Silencing TPX2 gene expression reduced the proliferation and migration(P<0.05),increased the percentage of G2 and S phase cell in the cell cycle(P<0.05),up-regulated the expression of P53,and down-regulated the expression of Ki-67,cyclin B2 and Aurora-A in HeLa cells (all P<0.05). CONCLUSION:Our results show that silencing TPX2 gene expression reduced the proliferation and migration in HeLa cells,which may be related to changes in cell cycle distribution,up-regulating the expression of P53,and down-regulate the expression of Ki-67,cyclin B2 and Aurora-A in HeLa cells.关键词
宫颈癌/TPX2基因/慢病毒/细胞增殖/细胞周期Key words
cervical cancer/TPX2 gene/lentivirus/cell proliferation/cell cycle分类
医药卫生引用本文复制引用
宋淑芳,常海平,杨彩容,赵荣伟,张龙,任杰,郑健..慢病毒介导的TPX2基因沉默对人宫颈癌HeLa细胞系增殖、迁移和细胞周期的影响及机制[J].癌变·畸变·突变,2017,29(5):329-334,339,7.基金项目
国家自然科学基金资助项目(81360385) （81360385）
