Actin bundling by dynamin 2 and cortactin is implicated in cell migration by stabilizing filopodia in human non-small cell lung carcinoma cells.
Actin bundling by dynamin 2 and cortactin is implicated in cell migration by stabilizing filopodia in human non-small cell lung carcinoma cells.
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DOI：
10.3892/ijo.2016.3592
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发表时间：
2016-09
影响因子：
5.2
通讯作者：
Takei K
中科院分区：
文献类型：
--
作者：
Yamada H;Takeda T;Michiue H;Abe T;Takei K
内吞蛋白发动蛋白参与形成肌动蛋白为基础的膜突起，如podosomes，伪足，和invadopodia，促进癌细胞迁移，侵袭和转移。然而，发动蛋白在癌细胞前缘形成肌动蛋白膜突起中的作用尚不清楚。在这项研究中，我们证明，普遍表达的发动蛋白2亚型促进细胞迁移的稳定F-肌动蛋白束丝状伪足的肺癌细胞系H1299。动力蛋白2的药理学抑制降低了细胞迁移和丝状伪足的形成。此外，免疫荧光和免疫电镜观察发现，血清刺激的H1299细胞的丝状伪足中，发动蛋白2和coronin主要共定位于沿着F-肌动蛋白束。敲低发动蛋白2或coronin抑制血清刺激的H1299细胞中丝状伪足的形成，伴随着F-肌动蛋白束的损失。表达野生型corpine拯救了发动蛋白2的点状定位和丝状伪足的形成。与F-肌动蛋白的孵育的发动蛋白2和corneumen诱导形成长而厚的肌动蛋白束，这些蛋白质共定位在F-肌动蛋白束。解聚试验表明，发动蛋白2和coronin增加了F-肌动蛋白束的稳定性。这些结果表明，发动蛋白2和corneumin参与细胞迁移，稳定F-肌动蛋白束在丝状伪足。总之，这些发现表明发动蛋白可能是一个可能的抗癌治疗的分子靶点。
The endocytic protein dynamin participates in the formation of actin-based membrane protrusions such as podosomes, pseudopodia, and invadopodia, which facilitate cancer cell migration, invasion, and metastasis. However, the role of dynamin in the formation of actin-based membrane protrusions at the leading edge of cancer cells is unclear. In this study, we demonstrate that the ubiquitously expressed dynamin 2 isoform facilitates cell migration by stabilizing F-actin bundles in filopodia of the lung cancer cell line H1299. Pharmacological inhibition of dynamin 2 decreased cell migration and filopodial formation. Furthermore, dynamin 2 and cortactin mostly colocalized along F-actin bundles in filopodia of serum-stimulated H1299 cells by immunofluorescent and immunoelectron microscopy. Knockdown of dynamin 2 or cortactin inhibited the formation of filopodia in serum-stimulated H1299 cells, concomitant with a loss of F-actin bundles. Expression of wild-type cortactin rescued the punctate-like localization of dynamin 2 and filopodial formation. The incubation of dynamin 2 and cortactin with F-actin induced the formation of long and thick actin bundles, with these proteins colocalizing at F-actin bundles. A depolymerization assay revealed that dynamin 2 and cortactin increased the stability of F-actin bundles. These results indicate that dynamin 2 and cortactin participate in cell migration by stabilizing F-actin bundles in filopodia. Taken together, these findings suggest that dynamin might be a possible molecular target for anticancer therapy.