Functional consequences of WNT3/Frizzled7-mediated signaling in non-transformed hepatic cells.
Functional consequences of WNT3/Frizzled7-mediated signaling in non-transformed hepatic cells.
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DOI：
10.1038/oncsis.2012.31
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发表时间：
2012-10-22
期刊：
影响因子：
6.2
通讯作者：
Kim, M.
中科院分区：
文献类型：
--
作者：
Nambotin, S. B.;Tomimaru, Y.;Merle, P.;Wands, J. R.;Kim, M.
我们之前已经证明，WNT3 和 Frizzled7 (FZD7) 的表达水平在肝细胞癌 (HCC) 中上调，并且它们直接相互作用以激活 HCC 细胞系中的经典 Wnt/β-catenin 通路。在本研究中，我们研究了未转化肝细胞中 WNT3 和 FZD7 表达水平的功能后果，以解决 WNT3/FZD7 介导的信号转导是否可能参与细胞转化的问题。稳定转染 WNT3 和 FZD7 后，在两种未转化的肝细胞来源的细胞系中通过蛋白质印迹、免疫染色和定量实时逆转录酶 PCR (qRT-PCR) 分析证实了 Wnt/β-catenin 通路的激活。测量了恶性表型的体外特征，包括软琼脂中的细胞增殖、迁移、侵袭和不依赖贴壁的生长。 WNT3和FZD7在两种细胞系中的稳定表达导致β-连环蛋白的细胞积累以及由该途径激活的下游靶基因的表达。在稳定表达WNT3/FZD7的克隆中，与未转化的对照细胞相比，肝细胞增殖、迁移、侵袭以及软琼脂集落形成均增强。表达 WNT3 和 FZD7 的细胞中上皮间质转化 (EMT) 因子 Twist、Snail 和 Vimentin 增加。然而，表达WNT3/FZD7的细胞在体内并未形成肿瘤。我们得出的结论是，WNT3/FZD7 经典途径的激活通过促进具有 EMT 特征的恶性表型的获得，在肝癌发生的早期阶段发挥作用。
We have previously demonstrated that WNT3 and Frizzled7 (FZD7) expression levelswere upregulated in hepatocellular carcinoma (HCC) and that they directly interact to activate the canonical Wnt/β–catenin pathway in HCC cell lines. In this study, we investigated the functional consequences of WNT3 and FZD7 expression levels in non-transformed hepatic cells to address the question of whether WNT3/FZD7-mediated signal transduction could be involved in cellular transformation. After stable transfection of WNT3 and FZD7, the activation of the Wnt/β–catenin pathway was confirmed by western blot, immunostaining and quantitative real-time reverse transcriptase–PCR (qRT–PCR) analysis in two non-transformed hepatocyte-derived cell lines. In vitro characteristics of the malignant phenotype were measured, including cell proliferation, migration, invasion and anchorage-independent growth in soft agar. Stable expression of WNT3 and FZD7 in the two cell lines led to cellular accumulation of β-catenin and expression of downstream target genes activated by this pathway. In the stable WNT3/FZD7-expressing clones, hepatic cell proliferation, migration, invasion as well as soft agar colony formation were enhanced compared with the non-transformed control cells. The epithelial–mesenchymal transition (EMT) factors, Twist, Snail and Vimentin, were increased in cells expressing WNT3 and FZD7. However, the WNT3/FZD7-expressing cells did not form tumors in vivo. We conclude that activation of the WNT3/FZD7 canonical pathway has a role in the early stages of hepatocarcinogenesis by promoting the acquisition of a malignant phenotype with features of EMT.
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影响因子：
13.5
作者：
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影响因子：
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影响因子：
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