NDRG2上调PTEN表达抑制乳腺癌细胞迁移的实验

目的利用具有相同遗传背景但不同转移能力的乳腺癌细胞模型MCF-7、LM-MCF-7以及MDA-MB-231细胞,研究NDRG2调控乳腺癌细胞迁移的作用及机制。方法利用RT-PCR和蛋白免疫印迹法检测MCF-7、LM-MCF-7以及MDA-MB-231细胞中NDRG2和PTEN mRNA和蛋白表达水平;构建pCMV-NDRG2和pCMV-PTEN表达载体以及设计合成NDRG2和PTEN siRNA,通过转染过表达或沉默NDRG2和PTEN表达,Transwell迁移实验检测转染后细胞迁移能力改变,Western blot检测NDRG2和PTEN表达调控关系。结果 MCF-7细胞中NDRG2和PT...

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Bibliographic Details
Published in肿瘤防治研究 Vol. 44; no. 1; pp. 5 - 10
Main Author 周晓雷 朱重悦 张世光 周志艳 李海潮 邹卫
Format Journal Article
LanguageChinese
Published 2017
Subjects
NDRG2
PTEN
乳腺肿瘤
细胞迁移
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Summary:目的利用具有相同遗传背景但不同转移能力的乳腺癌细胞模型MCF-7、LM-MCF-7以及MDA-MB-231细胞,研究NDRG2调控乳腺癌细胞迁移的作用及机制。方法利用RT-PCR和蛋白免疫印迹法检测MCF-7、LM-MCF-7以及MDA-MB-231细胞中NDRG2和PTEN mRNA和蛋白表达水平;构建pCMV-NDRG2和pCMV-PTEN表达载体以及设计合成NDRG2和PTEN siRNA,通过转染过表达或沉默NDRG2和PTEN表达,Transwell迁移实验检测转染后细胞迁移能力改变,Western blot检测NDRG2和PTEN表达调控关系。结果 MCF-7细胞中NDRG2和PTEN mRNA水平和蛋白表达水平显著高于LM-MCF-7和MDA-MB-231细胞（P〈0.05）;在LM-MCF-7和MDA-MB-231中上调NDRG2表达可降低细胞迁移能力至13.02%和26.33%（P〈0.01）;在MCF-7细胞中沉默NDRG2表达可增强细胞迁移能力至354.62%（P〈0.01）;蛋白免疫印迹检测显示PTEN表达受NDRG2调控;沉默（或过表达）NDRG2表达,同时过表达（或沉默）PTEN,MCF-7（或LM-MCF-7）细胞迁移能力未发生显著变化（P〉0.05）。结论乳腺癌细胞中,NDRG2的表达下调,导致PTEN表达下降,解除了对乳腺癌细胞迁移的抑制作用。NDRG2/PTEN信号途径的沉默是乳腺癌细胞获得高迁移能力的重要机制。
Bibliography:42-1241/R
Breast neoplasms; NDRG2; PTEN; Cell migration
Objective To investigate the effects of N-Myc downstream regulated gene 2（NDRG2） on the migration of human breast cancer cells by using two parallel cell lines MCF-7 and LM-MCF with different metastatic abilities. Methods The expression level of NDRG2 and PTEN in breast cancer cells were detected by RT-PCR and Western blot. The effects of overexpressed or down-regulated NDRG2 on the migration of breast cancer cells were investigated by Transwell migration test. The regulation mechanism between NDRG2 and PTEN on cell migration was detected via co-transfecting breast cancer cells with pCMV-NDRG2 and PTEN siRNA（or NDRG2 siRNA and pCMV-PTEN）. Results The m RNA and protein expression levels of NDRG2 and PTEN in MCF-7 cells were higher than those in LM-MCF-7/MDA-MB-231 cells（P〈0.05）. Overexpressed NDRG2 via transfecting LM-MCF-7 and MDA-MB-231 cells with pCMVNDRG2 could inhibit cell migration ability to 13.02% and 26.33%（P〈0.01） respectively. Downregulated NDRG2
ISSN:1000-8578