Jia Yongfeng, Wu Di, Yun Fen, Shi Lin, Luo Nianrong, Liu Zhiyue, Shi Yonghong, Sun Qinnuan, Jiang Lili, Wang Shiqi, Du Maolin
Department of Pathology, The First Affiliated Hospital of Inner Mongolia Medical University Huhhot, 010059, China.
Department of Pathology, Erdos City Central Hospital Erdos, 017000, China.
Int J Clin Exp Med. 2014 Apr 15;7(4):865-72. eCollection 2014.
Epithelial-mesenchymal transition (EMT) is associated with altered connection and junctions between cells and changes in abilities of invasion and migration. In this study, we investigated whether SK-BR-3 breast cancer cells induced to undergo EMT exhibit changes in morphological and invasion abilities after Transforming growth factor β1 (TGF-β1) treatment. Serum-deprived SK-BR-3 cells were treated with TGF-β1 (0, 10 ng/mL) for 24 h. The cells morphological changes were observed and imaged using inverted phase contrast microscope. Scratch experiment and invasion experiment were employed to detect changes of invasion ability, cell-flow experiment was used to assess cell cycle, immunohistochemistry technique was used to detect epithelial and mesenchymal markers after the crawling cells were fixed. Our research reveal that SK-BR-3 cells become larger and more messy, the elongated cells extend pseudopodia, the link of the cells became more loosely and cell gap widened after TGF-β1 treatment. SK-BR-3 cells showed faster growing and improved invasion abilities after TGF-β1 treatment, and reduced G1 phase cells proportion in the total number of cells after the conversion, in contrast the S phase cells accounted for the proportion of the total number of cells increased. These findings indicate that TGF-β1-induced EMT in breast cancer cells may be associated with major alterations in morphological and invasion abilities.
上皮-间质转化(EMT)与细胞间连接和黏附的改变以及侵袭和迁移能力的变化相关。在本研究中,我们调查了经诱导发生EMT的SK-BR-3乳腺癌细胞在转化生长因子β1(TGF-β1)处理后形态和侵袭能力是否发生变化。血清饥饿的SK-BR-3细胞用TGF-β1(0、10 ng/mL)处理24小时。使用倒置相差显微镜观察并拍摄细胞形态变化。采用划痕实验和侵袭实验检测侵袭能力的变化,使用细胞流式实验评估细胞周期,在爬行细胞固定后采用免疫组织化学技术检测上皮和间质标志物。我们的研究表明,TGF-β1处理后SK-BR-3细胞变大且更杂乱,伸长的细胞伸出伪足,细胞连接变得更松散且细胞间隙增宽。TGF-β1处理后SK-BR-3细胞生长加快且侵袭能力提高,转化后G1期细胞在细胞总数中的比例降低,相比之下S期细胞在细胞总数中的占比增加。这些发现表明,TGF-β1诱导的乳腺癌细胞EMT可能与形态和侵袭能力的主要改变有关。