解剖学报

解剖学报 ›› 2019, Vol. 50 ›› Issue (3): 304-309.doi: 10.16098/j.issn.0529-1356.2019.03.007

• 细胞和分子生物学 • 上一篇    下一篇

斑点型锌指结构蛋白在体外负性调控HepG2细胞的转移

梁少华 王志强 纪丕友*   

  1. 滨州医学院人体解剖学教研室，山东 烟台 264003
  • 收稿日期:2018-05-15 修回日期:2018-09-09 出版日期:2019-06-06 发布日期:2019-06-06
  • 通讯作者: 纪丕友 E-mail:jpy821101@163.com
  • 基金资助:
    山东省自然科学基金;山东省医药卫生科技发展计划项目;山东省医药卫生科技发展计划项目

Speckle-type POZ protein negatively regulating the metastasis of HepG2 cells in vitro

LIANG Shao-hua WANG Zhi-qiang JI Pi-you*   

  1. Department of Human Anatomy, Binzhou Medical University, Shandong Yantai 264003, China
  • Received:2018-05-15 Revised:2018-09-09 Online:2019-06-06 Published:2019-06-06
  • Contact: JI Pi-you E-mail:jpy821101@163.com

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摘要：

目的 探索斑点型锌指结构蛋白（SPOP）在体外对HepG2细胞迁移和侵袭的影响。 方法 利用质粒过表达和shRNA技术构建稳定过表达和敲低SPOP的稳定转染细胞，Western blotting检测SPOP过表达和敲低效果，然后利用体外Transwell实验检测SPOP对HepG2细胞的迁移和侵袭能力的影响，最后利用划痕实验再次检测SPOP对HepG2细胞迁移能力的影响。 结果 稳定转染HepG2中SPOP过表达和敲低效果明显；Transwell实验结果显示，过表达SPOP后HepG2细胞的迁移和侵袭能力明显下降，敲低后其迁移和侵袭能力明显上升。划痕实验结果显示，SPOP对HepG2细胞的迁移能力影响与Transwell实验一致。 结论 SPOP在体外可以负性调控HepG2细胞的转移能力。

关键词: 斑点型锌指结构蛋白, HepG2细胞, 转移, 免疫印迹法, 划痕实验

Abstract:

Objective To explore the effect of speckle type POZ protein (SPOP) on migration and invasion of HepG2 cells in vitro. Methods Stable gain- and loss- of SPOP expression HepG2 cell lines were constructed using plasmid overexpression and shRNA technique. Western blotting was used to detect the overexpression and knockdown effect of SPOP, and the effects of SPOP on the migration and invasion of HepG2 cells was examined by Transwell assays and wound-healing assay was also used to detect HepG2 cells’ migration in vitro. Results The effect of gain- and loss- of SPOP expression was obvious in constructed stable HepG2 cells. HepG2 cells’migration and invasion were significantly decreased by gain-SPOP expression, while the cells’migration and invasion were obviously enhanced in loss-SPOP expression HepG2 cells. Wound-healing assay exhibited that SPOP overexpression suppressed HepG2 cells’ migration and that downregulation of SPOP promoted the migration of HepG2 cells. Conclusion SPOP could negatively regulate HepG2 cells’ metastasis in vitro.

Key words: SPeckle-type POZ protein, HepG2 cell, Metastasis, Western blotting, Wound healing assay