Silencing of Twist Expression by RNA Interference Suppresses Epithelial-mesenchymal Transition, Invasion, and Metastasis of Ovarian Cancer
Silencing of Twist Expression by RNA Interference Suppresses Epithelial-mesenchymal Transition, Invasion, and Metastasis of Ovarian Cancer
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DOI：
10.7314/apjcp.2012.13.9.4435
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发表时间：
2012-01-01
影响因子：
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通讯作者：
Hou, Jian-Qing
中科院分区：
文献类型：
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作者：
Wang, Wen-Shuang;Yang, Xing-Sheng;Hou, Jian-Qing
目的：探讨Twist基因在卵巢癌上皮间质转化中的作用。研究方法：设计、构建表达靶向Twist的小干扰RNA（siRNA）的RNA干扰质粒（Twist siRNA载体），并转染人卵巢癌细胞系A2780。在荧光显微镜下评估转染效率。RT-PCR检测转染pGenesil Twist shRNA质粒后A2780细胞Twist mRNA表达的变化。MTT法和粘附实验检测Twist shRNA转染A2870细胞后细胞增殖和粘附能力的变化。用Western blotting分析Twist shRNA质粒转染A2780后E-cadherin和N-cadherin蛋白表达的变化。结果：成功构建重组质粒pGenesil-Twist shRNA。培养48 h后，80%的细胞表达高强度GFP荧光且稳定。转染Twist shRNA质粒后，Twist的表达明显下降（P0.05），而A2780的粘附能力明显下降（P
Purpose: This study aimed to explore the role of the Twist gene in the epithelial-mesenchymal transition of ovarian cancer. Methods: An RNA interference plasmid expressing a small interfering RNA (siRNA)-targeting Twist (Twist siRNA vector) was designed, constructed, and transfected into the human ovarian cancer cell line A2780. Transfection efficiency was assessed under a fluorescence microscope. Changes in the expression of Twist mRNA in A2780 after transfection with the pGenesil Twist shRNA plasmid were analyzed through RT-PCR. MTT assays and adhesion experiments were applied to determine changes in proliferation and adhesion ability of A2870 after transfection with the Twist shRNA plasmid. Changes in the expression of the E-cadherin and N-cadherin proteins in A2780 after transfection with the Twist shRNA plasmid were analyzed using Western blotting. Result: The restructuring plasmid pGenesil-Twist shRNA was constructed successfully. After 48 h of culture, 80% of the cells expressed high-intensity GFP fluorescence and stability. The expression of Twist decreased significantly after the transfection of the Twist shRNA plasmid (P0.05) but the adhesion ability of A2780 decreased dramatically (P