CAPZA1 modulates EMT by regulating actin cytoskeleton remodelling in hepatocellular carcinoma.
CAPZA1 modulates EMT by regulating actin cytoskeleton remodelling in hepatocellular carcinoma.
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CAPZA1 通过调节肝细胞癌肌动蛋白细胞骨架重塑来调节 EMT
DOI：
10.1186/s13046-016-0474-0
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发表时间：
2017-01-16
期刊：
影响因子：
--
通讯作者：
Zheng S
中科院分区：
文献类型：
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作者：
Huang D;Cao L;Zheng S
上皮-间质转化（epithelial-mesenchymal transition，EMT）引起细胞骨架重塑、基因表达和细胞表型的改变。在此过程中，肌动蛋白丝组装在维持肿瘤细胞的形态和运动中起着重要作用。加帽蛋白（Capping protein）是一种与肌动蛋白结合的蛋白质复合物，可调节肌动蛋白细胞骨架的重塑。CAPZA 1是该复合物的α1亚基，我们推测CAPZA 1通过调节肌动蛋白丝的组装来调节EMT，从而降低肝癌细胞的转移能力。采用Western blotting和qPCR检测肝癌细胞中CAPZA 1、EMT标志物和EMT转录因子的表达。采用Transwell迁移和侵袭实验观察肝癌细胞的迁移和侵袭。细胞计数试剂盒-8（CCK-8）检测肝癌细胞增殖情况。免疫沉淀法检测CAPZA 1与肌动蛋白丝的相互作用。结果CAPZA 1表达水平与原发性肝癌的生物学特性及患者预后呈负相关。CAPZA 1的表达与肝癌细胞的迁移和侵袭呈负相关。CAPZA 1的下调促进了肝癌细胞的迁移和侵袭。相反，CAPZA 1过表达显著抑制HCC细胞的迁移和侵袭。此外，CAPZA 1表达水平与EMT标志物E-钙粘蛋白、N-钙粘蛋白和波形蛋白的表达相关。此外，Snail 1和ZEB 1的表达与CAPZA 1的表达水平呈负相关。结论CAPZA 1通过调节肌动蛋白细胞骨架重塑抑制肝癌细胞EMT，从而降低肝癌细胞的转移能力。总之，我们的数据表明，CAPZA 1可能是一个有用的生物标志物，用于临床确定肝癌患者的预后。
BackgroundEpithelial-mesenchymal transition (EMT) elicits dramatic changes, including cytoskeleton remodelling as well as changes in gene expression and cellular phenotypes. During this process, actin filament assembly plays an important role in maintaining the morphology and movement of tumour cells. Capping protein, a protein complex referred to as CapZ, is an actin-binding complex that can regulate actin cytoskeleton remodelling. CAPZA1 is the α1 subunit of this complex, and we hypothesized that CAPZA1 regulates EMT through the regulation of actin filaments assembly, thus reducing the metastatic ability of hepatocellular carcinoma (HCC) cells.MethodsImmunohistochemistry was used to detect CAPZA1 expression in 129 HCC tissues. Western blotting and qPCR were used to detect CAPZA1, EMT markers and EMT transcription factors in HCC cells. Transwell migration and invasion assays were performed to observe the migration and invasion of HCC cells. Cell Counting Kit-8 (CCK-8) was used to detect the proliferation of HCC cells. Immunoprecipitation was used to detect the interaction between CAPZA1 and actin filaments. Finally, a small animal magnetic resonance imager (MRI) was used to observe metastases in HCC cell xenografts in the liver.ResultsCAPZA1 expression levels were negatively correlated with the biological characteristics of primary HCC and patient prognosis. CAPZA1 expression was negatively correlated with the migration and invasion of HCC cells. CAPZA1 down regulation promoted the migration and invasion of HCC cells. Conversely, CAPZA1 overexpression significantly inhibited the migration and invasion of HCC cells. Moreover, CAPZA1 expression levels were correlated with the expression of the EMT markers E-cadherin, N-cadherin and Vimentin. Furthermore, the expression of Snail1 and ZEB1 were negatively correlated with CAPZA1 expression levels. Similarly, CAPZA1 significantly inhibited intrahepatic metastases of HCC cells in an orthotopic transplantation tumour model.ConclusionsCAPZA1 inhibits EMT in HCC cells by regulating actin cytoskeleton remodelling, thereby reducing the metastatic ability of the cells. Together, our data suggest that CAPZA1 could be a useful biomarker for clinical determination of the prognosis of HCC patients.