Dual actin-bundling and protein kinase c-binding activities of fascin regulate carcinoma cell migration downstream of rac and contribute to metastasis
Dual actin-bundling and protein kinase c-binding activities of fascin regulate carcinoma cell migration downstream of rac and contribute to metastasis
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DOI：
10.1091/mbc.e07-02-0157
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发表时间：
2007-11-01
影响因子：
3.3
通讯作者：
Adams, Josephine C.
中科院分区：
文献类型：
--
作者：
Hashimoto, Yosuke;Parsons, Maddy;Adams, Josephine C.
由于细胞从原发肿瘤中迁移而导致的肿瘤复发是癌症治疗中的一个主要问题。人类癌症的免疫组织化学分析一致表明，肌动蛋白捆绑蛋白束蛋白的上调与临床侵袭性表型和不良预后相关。为了了解束蛋白的功能和机制贡献，我们在来自侵袭性原发肿瘤的人结肠癌细胞中进行了诱导短发夹RNA （shRNA）敲低束蛋白的实验。筋膜蛋白缺失导致丝状伪足数量减少，细胞突起形态改变，层粘连蛋白依赖rac的迁移减少，局灶粘连的周转减少，并且在体内减少异种移植物肿瘤的发展和转移。野生型绿色荧光蛋白-束蛋白或蛋白激酶C （PKC）磷酸化位点突变的束蛋白shrna敲低细胞的cDNA拯救表明，束蛋白的肌动蛋白结合和活跃的PKC结合活性都是丝状突起组织、rac依赖性迁移和肿瘤转移所必需的。因此，筋膜蛋白通过影响细胞迁移所需的多个亚细胞结构的双重途径参与肿瘤的迁移和转移。
Recurrence of carcinomas due to cells that migrate away from the primary tumor is a major problem in cancer treatment. Immunohistochemical analyses of human carcinomas have consistently correlated up-regulation of the actin-bundling protein fascin with a clinically aggressive phenotype and poor prognosis. To understand the functional and mechanistic contributions of fascin, we undertook inducible short hairpin RNA (shRNA) knockdown of fascin in human colon carcinoma cells derived from an aggressive primary tumor. Fascin-depletion led to decreased numbers of filopodia and altered morphology of cell protrusions, decreased Rac-dependent migration on laminin, decreased turnover of focal adhesions, and, in vivo, decreased xenograft tumor development and metastasis. cDNA rescue of fascin shRNA-knockdown cells with wild-type green fluorescent protein-fascin or fascins mutated at the protein kinase C (PKC) phosphorylation site revealed that both the actin-bundling and active PKC-binding activities of fascin are required for the organization of filopodial protrusions, Rac-dependent migration, and tumor metastasis. Thus, fascin contributes to carcinoma migration and metastasis through dual pathways that impact on multiple subcellular structures needed for cell migration.