慢病毒介导稳定敲低Smurf1细胞株的构建及对细胞迁移的影响
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协和青年基金资助;中央高校基本科研业务费专项资金资助(3332016077);中国医学科学院医学实验动物研究所基本科研业务费专项资助(2016ZX310033)


Establishment of two human tumor cell lines with lentivirus-mediated stably Smurf1 silencing and their effect on cell migration
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    目的 构建慢病毒介导稳定敲低Smurf1的HeLa和A549细胞株并检测敲低Smurf1细胞迁移的影响。方法 将包装好的Smurf1敲低慢病毒感染HeLa和A549细胞,7 d后进行嘌呤霉素抗性筛选阳性细胞,Western blot和qPCR检测敲低效果,并进行Transwell检测Smurf1敲低对细胞迁移的影响。结果 利用干扰慢病毒系统成功构建稳定敲低Smurf1的HeLa和A549细胞株,稳定敲低Smurf1抑制细胞的迁移速率。结论 敲低Smurf1抑制细胞迁移。

    Abstract:

    Objective To establish lentiviral expression vectors for Smurf1 silencing and assess the effects of Smurf1 silencing on cell migration. Methods HeLa and A549 cells were infected with lentiviral expression vectors for Smurf1 silencing respectively. After 7 days, the stable cell lines with Smurf1 silencing were obtained after puromycin-resistance screening, enrichment and expansion. The intracellular gene and protein levels of Smurf1 were detected by qPCR and western blot. Transwell assay was used to assess the effect of Smurf1 silencing on cell migration. Results The stable cell lines with Smurf1 silencing are constructed successfully. Silencing of Smurf1 down-regulated cell migration rate detected by Transwell assay. Conclusion Smurf1 promotes cell migration.

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韦荣飞,郭静,李梦媛,朱瑞敏,杨星九,高苒.慢病毒介导稳定敲低Smurf1细胞株的构建及对细胞迁移的影响[J].中国比较医学杂志,2017,27(4):46~51.

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  • 收稿日期:2016-11-26
  • 在线发布日期: 2017-04-28
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