Activation of Rho GTPase Cdc42 promotes adhesion and invasion in colorectal cancer cells.
Activation of Rho GTPase Cdc42 promotes adhesion and invasion in colorectal cancer cells.
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DOI：
10.12659/msmbr.883983
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发表时间：
2013-07-25
影响因子：
2.8
通讯作者：
Nan Qz
中科院分区：
文献类型：
--
作者：
Gao L;Bai L;Nan Qz
本研究旨在探讨活化的Rho GTPase细胞分裂控制蛋白42同源物（Cdc42）在结直肠癌细胞粘附、迁移和侵袭中的作用。组成活性形式的Cdc42 （GFP-Cdc42L61）或对照载体在结直肠癌细胞系SW480中过表达。通过免疫荧光染色监测活性Cdc42的定位，并通过附着实验、伤口愈合实验和体外Matrigel迁移实验检测活性Cdc42对细胞迁移和侵袭的影响。免疫荧光染色显示组成活性Cdc42主要定位于质膜。与转染对照载体的SW480细胞相比，在SW480细胞中过表达本构活性Cdc42可促进丝足形成和细胞拉伸，并显著增强细胞对包被板的粘附。伤口愈合实验显示，与对照细胞相比，表达活性Cdc42的SW480细胞的迁移能力显著增加。此外，Matrigel侵袭实验表明，活性Cdc42显著促进SW480细胞通过腔室的迁移。我们的研究结果表明，Rho GTPase Cdc42的活性可以极大地促进结直肠癌细胞SW480的扩散、迁移和侵袭，这可能有助于结直肠癌的转移。
The purpose of this study was to investigate the role of activated Rho GTPase cell division control protein 42 homolog (Cdc42) in colorectal cancer cell adhesion, migration, and invasion. The constitutively active form of Cdc42 (GFP-Cdc42L61) or control vector was overexpressed in the colorectal cancer cell line SW480. The localization of active Cdc42 was monitored by immunofluorescence staining, and the effects of active Cdc42 on cell migration and invasion were examined using an attachment assay, a wound healing assay, and a Matrigel migration assay in vitro. Immunofluorescence staining revealed that constitutively active Cdc42 predominately localized to the plasma membrane. Compared to SW480 cells transfected with the control vector, overexpression of constitutively active Cdc42 in SW480 cells promoted filopodia formation and cell stretch and dramatically enhanced cell adhesion to the coated plates. The wound healing assay revealed a significant increase of migration capability in SW480 cells expressing active Cdc42 compared to the control cells. Additionally, the Matrigel invasion assay demonstrated that active Cdc42 significantly promoted SW480 cell migration through the chamber. Our results suggest that active Rho GTPase Cdc42 can greatly enhance colorectal cancer cell SW480 to spread, migrate, and invade, which may contribute to colorectal cancer metastasis.