引用本文: |
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周奇,李婷,王国增,等.Twist shRNA对人宫颈癌细胞体外黏附、铺展及迁移能力的影响[J].同济大学学报(医学版),2011,32(5):23-26. [点击复制]
- ZHOU Qi,LI Ting,WANG Guo-zeng,et al.Effect of Twist short hairpin RNA on adhesion, spreading and migration of human cervical cancer HeLa cells in vitro[J].同济大学学报(医学版),2011,32(5):23-26. [点击复制]
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摘要: |
目的 通过RNA干扰抑制Twist基因在人宫颈癌HeLa细胞中的表达,观察Twist基因沉默对HeLa细胞体外黏附、铺展及迁移能力的影响。方法 根据shRNA设计原则,构建两种靶向Twist基因的短发夹RNA(shRNA)干扰质粒,稳定转染HeLa细胞。通过荧光定量PCR及Western印迹法检测HeLa细胞中Twist基因mRNA和蛋白的表达水平,利用黏附实验、铺展实验和划痕实验检测其对细胞黏附力和迁移力的影响。结果 成功构建的Twist shRNA真核表达载体稳定转染HeLa细胞后可显著降低Twist基因的表达。与对照组相比,Twist基因沉默组细胞的黏附、铺展及迁移能力明显下降(P<0.05)。结论 成功构建的Twist shRNA真核表达载体,能有效抑制HeLa细胞黏附、铺展及迁移能力。 |
关键词: 宫颈肿瘤 Twist RNA干扰 细胞黏附 细胞铺展 细胞迁移 |
DOI:10.3969/j.issn1008-0392.2011.05.006 |
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基金项目: |
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Effect of Twist short hairpin RNA on adhesion, spreading and migration of human cervical cancer HeLa cells in vitro |
ZHOU Qi,LI Ting,WANG Guo-zeng,ZHAO Jing,LIU Lan,WU Qian-yu |
(Dept.of Obstetrics and Gynecology,East Hospital,Tongji University,Shanghai 200120,China) |
Abstract: |
Objective To investigate the effect of shRNA of Twist gene on adhesion,spreading and migration of human cervical cancer HeLa cells.Methods Two kinds of shRNA vectors targeting to Twist gene were constructed and stably transfected into HeLa cells.The expressions of Twist gene and protein were detected by fluorescent quantitative PCR and Western blotting.The cell adhesion assay, cell spreading assay and cell migration assay were performed.Results The recombinant plasmids were successfully constructed and stably transfected into HeLa cells.The expression of Twist gene in HeLa cells was depressed significantly.The cell adhesion,spreading and migration activity were inhibited(P<0.05).Conclusion The shRNA expressing plasmids targeting to Twist gene can inhibit the adhesion,spreading and migration activity of HeLa cells in vitro. |
Key words: cervical cancer Twist RNA interference cell adhesion cell spreading cell migration |