Breast cancer-specific mutations in CK1epsilon inhibit Wnt/beta-catenin and activate the Wnt/Rac1/JNK and NFAT pathways to decrease cell adhesion and promote cell migration.
Breast cancer-specific mutations in CK1epsilon inhibit Wnt/beta-catenin and activate the Wnt/Rac1/JNK and NFAT pathways to decrease cell adhesion and promote cell migration.
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DOI：
10.1186/bcr2581
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Bryja V
中科院分区：
文献类型：
--
作者：
Foldynová-Trantírková S;Sekyrová P;Tmejová K;Brumovská E;Bernatík O;Blankenfeldt W;Krejcí P;Kozubík A;Dolezal T;Trantírek L;Bryja V
乳腺癌是女性中最常见的癌症之一。在乳腺癌中发现突变的基因之一是酪蛋白激酶1ε (CK1ε)。由于CK1ε是Wnt信号级联的关键调节因子，我们确定了这些CK1ε突变如何干扰Wnt通路并影响上皮性乳腺癌细胞系的行为。我们对各种突变进行了计算机建模，并分析了CK1ε突变体在体外和体内的激酶活性。此外，我们使用报告基因和小GTPase检测来确定CK1ε突变如何影响Wnt信号通路的不同分支。基于这些结果，我们对MCF7细胞进行了细胞粘附和细胞迁移实验，以证明CK1ε在这些过程中起着至关重要的作用。计算机模型和体内数据表明，在人类CK1ε n端叶中靠近乳腺癌点突变的Thr 44位点的自磷酸化参与了CK1ε活性的正向调节。我们的数据进一步表明，在哺乳动物细胞中，突变形式的CK1ε不能影响Dvl2的细胞内定位和磷酸化；我们能够证明CK1ε突变体不能增强dvl诱导的TCF/ lef介导的转录，CK1ε突变体在Wnt/β-catenin通路中起到功能丧失的作用，并且CK1ε突变体激活了非规范的Wnt/Rac-1和NFAT通路，类似于CK1的药物抑制剂。与这些发现一致，在乳腺癌来源的细胞系MCF7中，抑制CK1促进细胞迁移，降低细胞粘附和E-cadherin表达。综上所述，这些数据表明，在乳腺癌中发现的CK1ε突变可以抑制Wnt/β-catenin，并促进Wnt/Rac-1/JNK和Wnt/NFAT通路，从而通过影响细胞粘附和迁移来促进乳腺癌的发展。在分子机制方面，我们的数据表明，CK1ε n端叶的乳腺癌点突变与体外和体内CK1ε突变形式的磷酸化活性降低相关，干扰Thr 44的阳性自磷酸化。
Breast cancer is one of the most common types of cancer in women. One of the genes that were found mutated in breast cancer is casein kinase 1 epsilon (CK1ε). Because CK1ε is a crucial regulator of the Wnt signaling cascades, we determined how these CK1ε mutations interfere with the Wnt pathway and affect the behavior of epithelial breast cancer cell lines. We performed in silico modeling of various mutations and analyzed the kinase activity of the CK1ε mutants both in vitro and in vivo. Furthermore, we used reporter and small GTPase assays to identify how mutation of CK1ε affects different branches of the Wnt signaling pathway. Based on these results, we employed cell adhesion and cell migration assays in MCF7 cells to demonstrate a crucial role for CK1ε in these processes. In silico modeling and in vivo data showed that autophosphorylation at Thr 44, a site adjacent to the breast cancer point mutations in the N-terminal lobe of human CK1ε, is involved in positive regulation of the CK1ε activity. Our data further demonstrate that, in mammalian cells, mutated forms of CK1ε failed to affect the intracellular localization and phosphorylation of Dvl2; we were able to demonstrate that CK1ε mutants were unable to enhance Dvl-induced TCF/LEF-mediated transcription, that CK1ε mutants acted as loss-of-function in the Wnt/β-catenin pathway, and that CK1ε mutants activated the noncanonical Wnt/Rac-1 and NFAT pathways, similar to pharmacological inhibitors of CK1. In line with these findings, inhibition of CK1 promoted cell migration as well as decreased cell adhesion and E-cadherin expression in the breast cancer-derived cell line MCF7. In summary, these data suggest that the mutations of CK1ε found in breast cancer can suppress Wnt/β-catenin as well as promote the Wnt/Rac-1/JNK and Wnt/NFAT pathways, thus contributing to breast cancer development via effects on cell adhesion and migration. In terms of molecular mechanism, our data indicate that the breast cancer point mutations in the N-terminal lobe of CK1ε, which are correlated with decreased phosphorylation activities of mutated forms of CK1ε both in vitro and in vivo, interfere with positive autophosphorylation at Thr 44.
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影响因子：
5.3
作者：
Dejmek, Janna;Safholm, Annette;Leandersson, Karin
通讯作者：
Leandersson, Karin
DOI：
10.1073/pnas.060025397
发表时间：
2000-04-11
影响因子：
11.1
作者：
Lin, SY;Xia, WY;Hung, MC
通讯作者：
Hung, MC
影响因子：
7.7
作者：
Bryja, Vitezslav;Schambony, Alexandra;Schulte, Gunnar
通讯作者：
Schulte, Gunnar
影响因子：
21.3
作者：
Angers, S;Thorpe, CJ;Moon, RT
通讯作者：
Moon, RT
影响因子：
4.8
作者：
Lee, JS;Ishimoto, A;Yanagawa, S
通讯作者：
Yanagawa, S