Twist expression promotes migration and invasion in hepatocellular carcinoma.
Twist expression promotes migration and invasion in hepatocellular carcinoma.
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DOI：
10.1186/1471-2407-9-240
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发表时间：
2009-07-18
期刊：
影响因子：
3.8
通讯作者：
Yamamoto K
中科院分区：
文献类型：
--
作者：
Matsuo N;Shiraha H;Fujikawa T;Takaoka N;Ueda N;Tanaka S;Nishina S;Nakanishi Y;Uemura M;Takaki A;Nakamura S;Kobayashi Y;Nouso K;Yagi T;Yamamoto K
据报道，Twist是一种碱性螺旋-环-螺旋类转录因子，可以调节癌症转移。已知其诱导上皮-间质转化（EMT）。在这项研究中，我们评估了扭曲的表达及其对肝细胞癌（HCC）细胞迁移的影响。我们用免疫组织化学方法检测了20例肝细胞癌组织中Twist的表达，并通过RT-PCR和Western blot分析评估了Twist在肝癌细胞系中的表达。通过在扭转阴性HCC细胞系中引入扭转构建体来产生异位扭转表达。Twist siRNA阻断Twist阳性肝癌细胞系中内源性Twist表达。我们研究了EMT相关标志物，E-cadherin，Vimentin，和N-cadherin通过Western blot分析。MTT法检测细胞增殖情况，体外伤口愈合实验检测细胞迁移情况。我们使用免疫荧光黏着斑染色可视化粘着斑。我们在20个肿瘤样本中的7个中检测到强和中等扭曲表达，并且在无肿瘤切除边缘中没有发现显著的扭曲表达。此外，我们在HLE、HLF和SK-Hep 1细胞中检测到扭曲表达，但在PLC/RPF/5、HepG 2和Huh 7细胞中未检测到。异位扭转表达细胞表现出增强的细胞运动性，但扭转表达不影响细胞增殖。Twist表达诱导上皮-间充质转化以及相关的形态学变化。在异位扭转表达细胞中，粘着斑接触显著减少。Twist-siRNA处理的HLE、HLF和SK-Hep 1细胞表现出细胞迁移分别减少50%、40%和18%。Twist通过引起上皮-间质转化诱导肝癌细胞迁移。
Twist, a transcription factor of the basic helix-loop-helix class, is reported to regulate cancer metastasis. It is known to induce epithelial-mesenchymal transition (EMT). In this study, we evaluated the expression of twist and its effect on cell migration in hepatocellular carcinoma (HCC). We examined twist expression using immunohistochemistry in 20 tissue samples of hepatocellular carcinoma, and assessed twist expression in HCC cell lines by RT-PCR and Western blot analysis. Ectopic twist expression was created by introducing a twist construct in the twist-negative HCC cell lines. Endogenous twist expression was blocked by twist siRNA in the twist-positive HCC cell lines. We studied EMT related markers, E-cadherin, Vimentin, and N-cadherin by Western blot analysis. Cell proliferation was measured by MTT assay, and cell migration was measured by in vitro wound healing assay. We used immunofluorescent vinculin staining to visualize focal adhesion. We detected strong and intermediate twist expression in 7 of 20 tumor samples, and no significant twist expression was found in the tumor-free resection margins. In addition, we detected twist expression in HLE, HLF, and SK-Hep1 cells, but not in PLC/RPF/5, HepG2, and Huh7 cells. Ectopic twist-expressing cells demonstrated enhanced cell motility, but twist expression did not affect cell proliferation. Twist expression induced epithelial-mesenchymal transition together with related morphologic changes. Focal adhesion contact was reduced significantly in ectopic twist-expressing cells. Twist-siRNA-treated HLE, HLF, and SK-Hep1 cells demonstrated a reduction in cell migration by 50, 40 and 18%, respectively. Twist induces migratory effect on hepatocellular carcinoma by causing epithelial-mesenchymal transition.
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影响因子：
56.9
作者：
Raftopoulou, M;Etienne-Manneville, S;Hall, A
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