Zou Yuankang, Zhang Yimeng, Wang Ting, Wang Shan, Yang Angang, Jia Lintao, Wang Lei
Department of Biochemistry and Molecular Biology, Student Brigade, Fourth Military Medical University, Xi'an 710032, China.
Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2016 Feb;32(2):158-62.
To investigate the role of transmembrane prostate androgen-induced protein 1 (PMEPA1), an important gene downstream of transforming growth factor-β (TGF-β) signaling, in the process of breast cancer cell migration and epithelial-mesenchymal transition.
We treated MDA-MB-231 breast cancer cells with TGF-β and TGF-β inhibitor SB431542, and then detect the level of PMEPA1 using Western blotting. PMEPA1-specific siRNA was designed and its knockdown efficiency was tested by quantitative real-time PCR (qRT-PCR). After the expression of PMEPA1 in MDA-MB-231 cells was successfully silenced, the wound-healing assay and Transwell(TM) assay were used to investigate the effect of PMEPA1 silencing on the migration of MDA-MB-231 cells. Moreover, phalloidin was used to label the actin cytoskeleton of breast cancer cells to observe the effect of PMEPA1 silencing on cell morphology.
In breast cancer cells, PMEPA1 was upregulated by classical TGF-β/Smad signaling pathway. Silencing of PMEPA1 significantly inhibited the migration ability of MDA-MB-231 cells and promoted the process of mesenchymal-epithelial transition.
Over-expressed PMEPA1 can promote cell migration and maintain the mesenchymal-like morphology of breast cancer cells.
探讨跨膜前列腺雄激素诱导蛋白1（PMEPA1），一种转化生长因子-β（TGF-β）信号下游的重要基因，在乳腺癌细胞迁移和上皮-间质转化过程中的作用。
我们用TGF-β和TGF-β抑制剂SB431542处理MDA-MB-231乳腺癌细胞，然后用蛋白质印迹法检测PMEPA1的水平。设计PMEPA1特异性小干扰RNA（siRNA），并通过定量实时聚合酶链反应（qRT-PCR）检测其敲低效率。在成功沉默MDA-MB-231细胞中PMEPA1的表达后，采用划痕实验和Transwell实验研究PMEPA1沉默对MDA-MB-231细胞迁移的影响。此外，用鬼笔环肽标记乳腺癌细胞的肌动蛋白细胞骨架，以观察PMEPA1沉默对细胞形态的影响。
在乳腺癌细胞中，PMEPA1通过经典的TGF-β/Smad信号通路上调。PMEPA1沉默显著抑制MDA-MB-231细胞的迁移能力，并促进间质-上皮转化过程。
过表达的PMEPA1可促进细胞迁移并维持乳腺癌细胞的间质样形态。
