青岛大学医学院学报2009，Vol.45Issue(3)：297-300,302,5.DOI:10.3969/j.issn.1672-4488.2009.03.087
短发夹 RNA靶向抑制EGFR基因对人卵巢癌Skov-3细胞凋亡和细胞周期的影响
EFFECT OF EGFR GENE SILENCING ON CELL CYCLE AND APOPTOSIS OF OVARIAN CANCER CELL LINE SKOV-3
张红玲 1陈爱平 1杨蕊蕊1
作者信息
- 1. 青岛大学医学院附属医院妇科,山东,青岛,266003
- 折叠
摘要
Abstract
ObjectiveTo explore the effect of RNA interference (RNAi)-mediated inhibition on epidermal growth factor receptor (EGFR) gene expression in ovarian cancer cells. MethodsThe DNA sequence of EGFR gene was synthesized in vitro and cloned into pSilencer 2.1-U6neo plasmid vector to construct the shRNA expression vector. After the constructed vector was transfected into human ovarian cancer Skov-3 cells using lipofectamine 2000, semi-quantitative RT-PCR and immunocytochemistry assay were conducted to compare the changes in the expression levels of EGFR between before and after transfection. Moreover, cells were stained with Annexin V/PI and PI and then siRNA-induced apoptosis and cell cycle analyzed. ResultsSiRNA eukaryotic expression vector pSilencer-EGFR was successfully constructed; the siRNA targeted against EGFR could effectively inhibit EGFR gene expression in Skov-3 cells; the results of flow cytometric analysis indicated that the apoptosis rate of Skov-3 cells increased significantly and an obvious cell cycle arrest at G0/G1 phase was induced.Conclusion RNAi-mediated silencing of EGFR gene expression can induce cell apoptosis and cell cycle arrest at G0/G1 phase. Thus, RNAi can be used as an effective tool to explore the mechanism underlying the development and progression of ovarian cancer as well as conduct gene therapy of this disease.关键词
卵巢肿瘤/受体,表皮生长因子/RNA干扰/细胞凋亡Key words
Ovarian neoplams/ Receptor, epidermal growth factor/ RNA interference/ Apoptosis分类
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张红玲,陈爱平,杨蕊蕊..短发夹 RNA靶向抑制EGFR基因对人卵巢癌Skov-3细胞凋亡和细胞周期的影响[J].青岛大学医学院学报,2009,45(3):297-300,302,5.
