Damnacanthal, an effective inhibitor of LIM-kinase, inhibits cell migration and invasion.
Damnacanthal, an effective inhibitor of LIM-kinase, inhibits cell migration and invasion.
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DOI：
10.1091/mbc.e13-09-0540
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发表时间：
2014-03
影响因子：
3.3
通讯作者：
Mizuno K
中科院分区：
文献类型：
--
作者：
Ohashi K;Sampei K;Nakagawa M;Uchiumi N;Amanuma T;Aiba S;Oikawa M;Mizuno K
丹那坎塔尔是一种有效的lim激酶抑制剂。抑制t细胞的趋化性和乳腺癌细胞在培养中的迁移和侵袭，以及半抗原诱导的小鼠耳表皮朗格汉斯细胞的迁移。Damnacanthal是研究lim激酶细胞和生理功能的有用工具。limk激酶（limk）通过磷酸化和失活cofilin，在多种细胞活动中发挥重要作用，包括迁移、分裂和形态发生。利用双分子荧光互补法检测肌动蛋白- cofilin相互作用，我们筛选了LIMK1抑制剂，并鉴定出两种有效的抑制剂，damnacanthal （Dam）和MO-26（吡唑啉嘧啶衍生物）。这些化合物已经被证明可以抑制Lck，一种Src家族酪氨酸激酶。然而，体外激酶实验显示，Dam比Lck更有效地抑制LIMK1。Dam抑制limk1诱导的cofilin磷酸化和N1E-115细胞板足肌动蛋白逆行流动的减速。Dam损害了cxcl12诱导的Jurkat T淋巴细胞和Jurkat来源的lck缺陷JCaM1.6细胞的趋化迁移，也抑制了血清诱导的MDA-MB-231乳腺癌细胞的迁移和侵袭。这些结果表明，Dam主要通过抑制LIMK激酶活性来抑制细胞迁移和侵袭。局部应用Dam还能抑制半抗原诱导的小鼠耳表皮朗格汉斯细胞的迁移。Dam为研究limk的细胞和生理功能提供了有用的工具，并为开发针对limk相关疾病的药物提供了希望。本研究中使用的双分子荧光互补分析系统将为筛选各种蛋白激酶抑制剂提供一种有用的方法。
Damnacanthal is identified as an effective inhibitor of LIM-kinase. It inhibits chemotaxis of T-cells and migration and invasion of breast carcinoma cells in culture and hapten-induced migration of epidermal Langerhans cells in mouse ears. Damnacanthal is a useful tool for investigating the cellular and physiological functions of LIM-kinase. LIM-kinases (LIMKs) play crucial roles in various cell activities, including migration, division, and morphogenesis, by phosphorylating and inactivating cofilin. Using a bimolecular fluorescence complementation assay to detect the actin–cofilin interaction, we screened LIMK1 inhibitors and identified two effective inhibitors, damnacanthal (Dam) and MO-26 (a pyrazolopyrimidine derivative). These compounds have already been shown to inhibit Lck, a Src family tyrosine kinase. However, in vitro kinase assays revealed that Dam inhibited LIMK1 more effectively than Lck. Dam suppressed LIMK1-induced cofilin phosphorylation and deceleration of actin retrograde flow in lamellipodia in N1E-115 cells. Dam impaired CXCL12-induced chemotactic migration of Jurkat T lymphocytes and Jurkat-derived, Lck-deficient JCaM1.6 cells and also inhibited serum-induced migration and invasion of MDA-MB-231 breast carcinoma cells. These results suggest that Dam has the potential to suppress cell migration and invasion primarily through the inhibition of LIMK kinase activity. Topical application of Dam also suppressed hapten-induced migration of epidermal Langerhans cells in mouse ears. Dam provides a useful tool for investigating cellular and physiological functions of LIMKs and holds promise for the development of agents against LIMK-related diseases. The bimolecular fluorescence complementation assay system used in this study will provide a useful method to screen for inhibitors of various protein kinases.