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

doi:10.3969/j.issn.1004-616x.2017.05.002

Abstract

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.

Key words: cervical cancer, TPX2 gene, lentivirus, cell proliferation, cell cycle

CLC Number:

R737.3

SONG Shufang, CHANG Haiping, YANG Cairong, ZHAO Rongwei, ZHANG Long, REN Jie, ZHENG Jian. Effect and mechanism of lentivirus-mediated silencing of TPX2 gene expression on proliferation, migration and cell cycle of HeLa cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(5): 329-334,339.

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Effect and mechanism of lentivirus-mediated silencing of TPX2 gene expression on proliferation, migration and cell cycle of HeLa cells

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