| 摘要: |
| 目的探讨siRNA沉默Msi1基因表达对人宫颈癌Hela细胞增殖、迁移及侵袭的影响。方法针对Msi1基因mRNA序列设计合成siRNA，转染人宫颈癌Hela细胞；实验分Msi1siRNA组、空白对照组、脂质体组及阴性对照组；通过RT-PCR法检测Msi1基因在mRNA水平的表达；分别利用四甲基偶氮唑盐（MTT）比色试验、群体倍增时间及平皿克隆实验分析干扰Msi1基因对人宫颈癌Hela细胞的生长抑制效应；通过细胞划痕实验观察干扰Msi1对细胞迁移能力的影响；利用Transwell小室实验观察干扰Msi1对细胞侵袭能力的影响。结果Msi1siRNA能有效抑制人宫颈癌Hela细胞中Msi1基因的表达；Msi1siRNA转染Hela细胞24、48、72h后，与其他各组比较，Msi1siRNA组Hela细胞的生长、增殖速度明显减缓（均P＜0.05）。空白对照组Hela细胞群体倍增时间为（20.18±0.01）h，而干扰Msi1基因处理后的Hela细胞，群体倍增时间延长至（35.68±0.04）h（P＜0.05）；同时平皿克隆实验结果发现，Msi1siRNA处理后的Hela细胞与其他各组相比，Hela细胞生长明显减慢（均P＜0.05）；细胞划痕实验和Transwell小室实验分别显示处理后的Hela细胞其迁移和侵袭能力均明显下降，与其他各组相比差异均有统计学意义（均P＜0.05）。结论沉默Msi1基因表达对人宫颈癌Hela细胞的增殖、迁移及侵袭有负性调节作用。 |
| 关键词: 宫颈癌 Msi1 siRNA 增殖 迁移 侵袭 |
| DOI：10.12056/j.issn.1006-2785.2018.40.12.2017-2476 |
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| Effect of Msi1 siRNA on proliferation, migration and invasion of Hela cells |
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| Abstract: |
| Objective To investigate the effect of silencing Msi1 gene with siRNA on proliferation, migration and invasion of Hela cells. Methods Msi1-specific siRNA was designed and synthesized, and the small interfering RNA was transfected into Hela cells to knockdown the Msi1 expression. The Hela cells divided into Msi1 siRNA group, blank control group, liposome group and negative control group. mRNA levels of Msi1 was detected by RT-PCR; MTT assay, cell average doubling time analysis and plate cloning experiment were employed to confirm Msi1 siRNA-induced cell growth inhibition in Hela cells. Cell migration and invasion were respectively detected by wound healing assay and Transwell assay. Results RNA interference efficiently suppressed the Msi1 expression in Hela cells. After transfection with Msi1 siRNA for 24, 48 and 72h, proliferation rate of Hela cells
was reduced(all P＜0.05) , the average doubling time retarded from (20.18±0.01)h of normal cultured Hela cells to (35.68±0.04)h
of Msi1 siRNA transfected Hela cells (P＜0.05); the plate cloning assay showed that the growth rate of Msi1 siRNA group was lower than other groups(all P＜0.05); wound healing assay and Transwell assay showed that the migration and invasion abilities of Hela cells were significantly reduced after Msi1 gene knockdowned (all P＜0.05). Conclusion Downregulation of Msi1 gene expression results in significant inhibition of proliferation, migration and invasion in Hela cells. |
| Key words: Uterine cervix cancer Msi1 siRNA Proliferation Migration Invasion |
