BVES Inhibition Triggers Epithelial-Mesenchymal Transition in Human Hepatocellular Carcinoma
BVES Inhibition Triggers Epithelial-Mesenchymal Transition in Human Hepatocellular Carcinoma
复制标题
BVES 抑制触发人肝细胞癌的上皮-间质转化
DOI：
10.1007/s10620-013-2992-3
复制
发表时间：
2014-05-01
影响因子：
3.1
通讯作者：
Yan, Wei
中科院分区：
文献类型：
--
作者：
Han, Ping;Fu, Yu;Yan, Wei
背景/目的转移导致肝细胞癌 (HCC) 的预后不良。然而，原发性 HCC 细胞发展成转移表型的机制尚不清楚。在本研究中，我们着手阐明血管心外膜物质（BVES）这种调节紧密连接形成的新型粘附分子如何介导人HCC细胞的侵袭和转移。方法采用qRT-PCR、蛋白质印迹和IHC来检测BVES在HCC样本和HCC细胞系中的表达。合成了针对人 BVES 的小干扰 RNA (siRNA)，并用于转染 Huh7 细胞。然后，通过qRT-PCR和Western blot检测干扰效率以及间质标记物波形蛋白和上皮标记物E-钙粘蛋白的表达。使用 TRITC 缀合的鬼笔环肽检测 F-肌动蛋白细胞骨架。抑制BVES后，通过伤口愈合实验和transwell实验分析Huh7细胞的迁移和侵袭能力。结果BVES在人HCC组织和高转移潜能HCC细胞系中表达下调。 BVES 抑制后，Huh7 细胞表现出一些形态学变化，包括细胞骨架重排和连接破坏。细胞迁移和侵袭随着波形蛋白、IL-6、MMP2、MMP9表达的增加和E-钙粘蛋白表达的减少而增加。最后，我们发现BVES敲低细胞中上皮间质转化（EMT）转录因子Snail1和Twist1的表达显着增加。结论我们的结果表明HCC中BVES的下调诱导EMT，从而促进HCC细胞的侵袭和转移。
Background/AimMetastasis contributes to the poor prognosis of hepatocellular carcinoma (HCC). However, the mechanism through which a primary HCC cell develops into a metastatic phenotype is not well understood. In this study, we set out to elucidate how blood vessel epicardial substance (BVES), a novel adhesion molecule regulating tight junction formation, mediates invasion and metastasis in human HCC cells.MethodsqRT-PCR, western blot and IHC were used to detect the expression of BVES in HCC samples and HCC cell lines. Small interfering RNAs (siRNAs) against human BVES were synthesized and used to transfect Huh7 cells. Then, the interference efficiency and the expression of mesenchymal marker vimentin and epithelial marker E-cadherin were measured by qRT-PCR and western blot. F-actin cytoskeleton was detected using TRITC-conjugated phalloidin. After inhibition of BVES, wound healing experiment and transwell assay were used to analyze the migratory and invasive ability of Huh7 cells.ResultsBVES was down-regulated in human HCC tissues and HCC cell lines with high metastatic potential. After BVES inhibition, Huh7 cells exhibited some morphological changes including cytoskeleton rearrangement and junctional disruption. Cell migration and invasion were increased concomitant with increased expression of vimentin, IL-6, MMP2, MMP9 and decreased expression of E-cadherin. Finally, we found the expression of epithelial-mesenchymal transition (EMT) transcription factors Snail1 and Twist1 was significantly increased in BVES knockdown cells.ConclusionsOur results suggest that down-regulation of BVES in HCC induces EMT, thus promoting invasion and metastasis in HCC cells.