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目的探究转化生长因子β(TGF-β)信号下游重要靶分子跨膜前列腺雄激素诱导蛋白1(PMEPA1)对乳腺癌细胞运动迁移能力的影响及其在上皮间质转化(EMT)过程中的作用。方法用TGF-β刺激MDA-MB-231乳腺癌细胞,并用小分子抑制剂SB431542抑制TGF-β信号通路后,采用Western blot法检测PMEPA1的表达水平;设计针对PMEPA1分子的特异性小干扰RNA(siRNA),采用实时定量PCR检测干涉效率;干涉MDA-MB-231细胞中PMEPA1的表达后,采用划痕实验和TranswellTM实验检测PMEPA1基因沉默对乳腺癌细胞迁移能力的影响;采用鬼笔环肽对细胞骨架进行标记,观察PMEPA1基因沉默后MDAMB-231乳腺癌细胞的形态变化。结果乳腺癌细胞中,PMEPA1分子受经典TGF-β/Smad信号途径的作用发生显著上调;沉默PMEPA1的表达能显著抑制乳腺癌细胞MDA-MB-231的迁移能力,并使细胞形态由间质样向上皮样转变。结论 PMEPA1促进乳腺癌细胞迁移并维持乳腺癌细胞的间质样特性。
Objective To investigate the effect of transmembrane prostate androgen receptor 1 (PMEPA1), an important downstream target of transforming growth factor β (TGF-β) signaling, on the motility and migration of breast cancer cells and its role in epithelial-mesenchymal transition (EMT) . Methods After stimulating MDA-MB-231 breast cancer cells with TGF-β, the expression of PMEPA1 was detected by Western blot after inhibiting the TGF-β signaling pathway with small molecule inhibitor SB431542. Specific siRNA targeting PMEPA1 was designed siRNA). The interference efficiency of PMEPA1 was detected by real-time quantitative PCR. After scratching the expression of PMEPA1 in MDA-MB-231 cells, scratch test and TranswellTM assay were used to detect the effect of PMEPA1 silencing on the migration ability of breast cancer cells. The cytoskeleton was labeled to observe the morphological changes of MDAMB-231 breast cancer cells after PMEPA1 gene silencing. Results The expression of PMEPA1 in breast cancer cells was significantly up-regulated by the classical TGF-β / Smad signaling pathway. The silencing of PMEPA1 significantly inhibited the migration of breast cancer cells MDA-MB-231, To epithelial-like change. Conclusion PMEPA1 can promote the migration of breast cancer cells and maintain the interstitial characteristics of breast cancer cells.