基础医学与临床 ›› 2018, Vol. 38 ›› Issue (6): 771-775.

• 研究论文 • 上一篇    下一篇

Twist促进人三阴性乳腺癌细胞迁移和侵袭

张达容1,王维莲2,吴立翔1,柳满然3   

  1. 1. 重庆大学附属肿瘤医院、重庆市肿瘤研究所、重庆市肿瘤医院
    2. 武警重庆市总队医院
    3. 重庆医科大学
  • 收稿日期:2018-01-29 修回日期:2018-04-21 出版日期:2018-06-05 发布日期:2018-05-25
  • 通讯作者: 柳满然 E-mail:mliu-hncq@hotmail.com
  • 基金资助:
    Twist调控乳腺上皮细胞EMT转化及肿瘤干细胞样细胞特征的机制研究

Twist promotes cell migration and invasion in human breast cancer cell line Hs578T

  • Received:2018-01-29 Revised:2018-04-21 Online:2018-06-05 Published:2018-05-25

摘要: 目的 研究Twist基因沉默对人三阴性乳腺癌Hs578T细胞迁移和侵袭的影响及其潜在机制。方法 构建靶向Twist基因的慢病毒载体(shTwist)及其阴性对照病毒(shNC),转染人三阴性乳腺癌Hs578T细胞,建立Twist基因沉默的稳定细胞系;实验设对照组(blank)、阴性对照组(shNC)以及Twist基因沉默组(shTwist);经嘌呤霉素筛选后,倒置荧光显微镜观察各组细胞的荧光表达和感染效率,用RT-qPCR和Western blot检测Twist mRNA和蛋白的表达水平;Transwell迁移和侵袭实验分别检测细胞的迁移和侵袭能力;Western blot进一步检测Twist下游p-AKT、AKT、p-ERK1/2和ERK1/2蛋白水平。结果 成功构建Twist基因稳定沉默人三阴性乳腺癌HS578T细胞系;与blank组和shNC组细胞相比,shTwist组细胞迁移和侵袭能力明显减弱(p<0.05, p<0.05);Twist下游p-AKT和p-ERK1/2蛋白水平显著下调(p<0.05, p<0.05)。结论 在人三阴性乳腺癌Hs578T细胞中,Twist可能通过AKT和ERK信号通路促进细胞迁移和侵袭。

关键词: Twist, 乳腺癌, 细胞迁移, 细胞侵袭

Abstract: Objective To study the effects of Twist gene silencing on human triple-negative breast cancer Hs578T cell migration and invasion and its potential mechanisms. Methods The lentiviral vectors with Twist gene-targeted specific shRNA and the negative control were constructed, then transfected into the human triple-negative breast cancer cell line Hs578T to establish the stable cell lines of Twist gene silencing. The blank group (blank), negative control group (shNC) and Twist gene silencing group (shTwist) were set up. After screening by puromycin, the fluorescence expression and the infection efficiency of each group cells were observed by inverted fluorescence microscope. The expression of Twsit mRNA and protein level was detected by quantitative real-time PCR and Western blot. Transwell migration and invasion experiments were performed to measure cell migration and invasion abilities, respectively. Western blot was used to detect the levels of p-AKT, AKT, p-ERK1/2 and ERK1/2 proteins. Results Twist gene was successfully knockdown in human triple-negative breast cancer cell line Hs578T. Compared with the blank group and shNC group cells, the cell migration and invasion ability of the shTwist group cells were decreased significantly (p<0.05, p<0.05), and the expression of Twist downstream p-AKT and p-ERK1/2 proteins were also reduced notably in the shTwist group cells (p<0.05, p<0.05). Conclusions Twist promotes cell migration and invasion via AKT/ERK signaling axis in human triple-negative breast cancer cell line Hs578T.

Key words: Twist, breast cancer, cell migration, cell invasion