NDRG2上调PTEN表达抑制乳腺癌细胞迁移的实验

发布时间：2018-03-21 00:54

本文选题：乳腺肿瘤　切入点：NDRG　出处：《肿瘤防治研究》2017年01期 　论文类型：期刊论文

【摘要】：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(P0.05);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%(P0.01);在MCF-7细胞中沉默NDRG2表达可增强细胞迁移能力至354.62%(P0.01);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(P0.05)。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。
[Abstract]:Objective to use breast cancer cell model MCF-7 LM-MCF-7 and MDA-MB-231 cells with the same genetic background but different metastatic ability. Methods the expression levels of NDRG2 and PTEN mRNA and protein in MCF-7 LM-MCF-7 and MDA-MB-231 cells were detected by RT-PCR and Western blot. The expression vectors of pCMV-NDRG2 and pCMV-PTEN were constructed, and NDRG2 and PTEN siRNAs were designed and synthesized. After transfection or silencing of NDRG2 and PTEN expression, Transwell migration assay was used to detect the ability of cell migration after transfection. Western blot was used to detect the regulatory relationship between NDRG2 and PTEN expression. The levels of NDRG2 and PTEN mRNA and protein expression were significantly higher in the cells than in LM-MCF-7 and MDA-MB-231 cells, the expression of NDRG2 in LM-MCF-7 and MDA-MB-231 decreased to 13.02% and 26.33% P0.01G, and the silencing of NDRG2 expression enhanced the migration ability to 354.62% (P0.01) in MCF-7 cells. Western blot analysis showed that the expression of PTEN was regulated by NDRG2. The expression of NdRG2 was silenced (or overexpressed) and the migration ability of MCF-7 (or LM-MCF-7) cells did not change significantly. Conclusion the expression of NDRG2 in breast cancer cells is down-regulated, resulting in the decrease of PTEN expression. The silencing of NDRG _ 2 / PTEN signaling pathway is an important mechanism for breast cancer cells to obtain high migration ability.
【作者单位】：河北科技大学生物制造公共实验中心;
【基金】：河北省高等学校科学研究项目(QN2016020) 2015年度河北科技大学五大平台开放基金课题河北科技大学博士科研启动基金课题
【分类号】：R737.9

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