Transient but not stable ZEB1 knockdown dramatically inhibits growth of malignant pleural mesothelioma cells.
Transient but not stable ZEB1 knockdown dramatically inhibits growth of malignant pleural mesothelioma cells.
复制标题
瞬态但不稳定的Zeb1敲低会大大抑制恶性胸膜间皮瘤细胞的生长。
DOI：
10.1245/s10434-011-2142-0
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发表时间：
2012-07
影响因子：
3.7
通讯作者：
Hasegawa Y
中科院分区：
文献类型：
--
作者：
Horio M;Sato M;Takeyama Y;Elshazley M;Yamashita R;Hase T;Yoshida K;Usami N;Yokoi K;Sekido Y;Kondo M;Toyokuni S;Gazdar AF;Minna JD;Hasegawa Y
ZEB 1是一种主要的上皮-间充质转化基因，在恶性胸膜间皮瘤（MPM）中的作用尚不清楚。采用实时定量聚合酶链反应和Western blot检测18个MPM细胞系和1个正常胸膜间皮瘤细胞系MeT-5A中ZEB 1、E-cadherin、vimentin和EpCAM的表达。进行RNA干扰介导的ZEB 1和EpCAM的瞬时和/或稳定敲低。用TORAY-3D基因芯片进行微阵列表达分析。通过比色增殖和集落形成试验评价生长。进行荧光素酶报告基因测定以评估ZEB 1敲低对EpCAM启动子活性的影响。大多数MPM细胞系表现出间充质表型，并表达ZEB 1。瞬时ZEB 1敲低抑制了研究的所有四种细胞系（ACC-MESO-1、H2052、Y-MESO-8A、Y-MESO-29）的生长，而稳定的ZEB 1敲低仅抑制了Y-MESO-29的生长。全基因组基因表达分析显示，EpCAM是ACC-MESO-1中通过瞬时和稳定ZEB 1敲除最显著上调的基因，在稳定敲除中上调更显著。我们假设EpCAM上调抵消了ACC-MESO-1中稳定的ZEB 1敲低诱导的生长抑制。瞬时EpCAM敲除显著抑制表达shZEB 1的ACC-MESO-1细胞的生长，但仅适度抑制表达shGFP的细胞的生长，支持我们的假设。荧光素酶报告基因测定显示ZEB 1敲低导致EpCAM启动子活性增加。EpCAM在表达shZEB 1的Y-MESO-29中也被上调，但这种EpCAM上调不抵消ZEB 1敲低诱导的生长抑制，表明EpCAM的抵消作用可能是细胞环境依赖性的。RNA干扰介导的ZEB 1敲低可能是MPM的一种有前途的治疗策略，但必须考虑长期ZEB 1敲低的生长抑制作用减弱的可能性，这可能是由于EpCAM上调和/或ZEB 1敲低导致的其他基因表达变化。
The role of ZEB1, a master epithelial-tomesenchymal transition gene, in malignant pleural mesothelioma (MPM) is unclear. The expression of ZEB1, E-cadherin, vimentin, and epithelial cell adhesion molecule (EpCAM) in 18 MPM cell lines and a normal pleural mesothelial cell line MeT-5A was determined by quantitative real-time polymerase chain reaction and Western blot testing. RNA interference–mediated transient and/or stable knockdown of ZEB1 and EpCAM was performed. Microarray expression analysis was performed with a TORAY-3D gene chip. Growth was evaluated by colorimetric proliferation and colony formation assays. Luciferase reporter assay was performed to access the effects of ZEB1 knockdown on EpCAM promoter activity. Most MPM cell lines exhibited mesenchymal phenotype and expressed ZEB1. Transient ZEB1 knockdown suppressed growth in all four cell lines studied (ACC-MESO-1, H2052, Y-MESO-8A, Y-MESO-29) while stable ZEB1 knockdown suppressed growth only in Y-MESO-29. Genome-wide gene expression analysis revealed that EpCAM was the most prominently up-regulated gene by both transient and stable ZEB1 knockdown in ACC-MESO-1, with more marked up-regulation in stable knockdown. We hypothesized that EpCAM up-regulation counteracts the stable ZEB1 knockdown-induced growth inhibition in ACC-MESO-1. Transient EpCAM knockdown suppressed growth dramatically in ACC-MESO-1 cells expressing shZEB1 but only modestly in those expressing shGFP, supporting our hypothesis. Luciferase reporter assay showed that ZEB1 knockdown resulted in increased EpCAM promoter activity. EpCAM was also up-regulated in Y-MESO-29 expressing shZEB1, but this EpCAM up-regulation did not counteract ZEB1 knockdown-induced growth suppression, suggesting that the counteracting effects of EpCAM may be cellular context dependent. RNA interference-mediated ZEB1 knockdown may be a promising therapeutic strategy for MPM, but one has to consider the possibility of diminished growth inhibitory effects of long-term ZEB1 knockdown, possibly as a result of EpCAM up-regulation and/or other gene expression changes resulting from ZEB1 knockdown.
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影响因子：
11.2
作者：
Spaderna, Simone;Schmalhofer, Otto;Brabletz, Thomas
通讯作者：
Brabletz, Thomas
影响因子：
11.2
作者：
Osta, WA;Chen, Y;Gillanders, WK
通讯作者：
Gillanders, WK
影响因子：
8
作者：
Eger, A;Aigner, K;Foisner, R
通讯作者：
Foisner, R
影响因子：
9.7
作者：
Takeyama, Yoshihiro;Sato, Mitsuo;Horio, Mihoko;Hase, Tetsunari;Yoshida, Kenya;Yokoyama, Toshihiko;Nakashima, Harunori;Hashimoto, Naozumi;Sekido, Yoshitaka;Gazdar, Adi F.;Minna, John D.;Kondo, Masashi;Hasegawa, Yoshinori
通讯作者：
Hasegawa, Yoshinori
影响因子：
11.2
作者：
Sato, M;Vaughan, MB;Minna, JD
通讯作者：
Minna, JD