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Activation of Rho GTPase Cdc42 promotes adhesion and invasion in colorectal cancer cells

机译:Rho GTPase Cdc42的激活促进结直肠癌细胞的粘附和侵袭

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Background 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. Material and Methods 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. Results 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. Conclusions 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.
机译:背景本研究的目的是研究活化的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。材料和方法Cdc42的组成型活性形式(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,并在体外使用附着试验,伤口愈合试验和基质胶迁移试验检查活性Cdc42对细胞迁移和侵袭的影响。结果免疫荧光染色显示,组成型活性Cdc42主要定位于质膜。与用对照载体转染的SW480细胞相比,SW480细胞中组成型活性Cdc42的过表达促进丝状伪足的形成和细胞舒张,并显着增强细胞对包被平板的粘附。伤口愈合试验显示,与对照细胞相比,表达活性Cdc42的SW480细胞的迁移能力显着提高。此外,Matrigel侵袭试验证明活性Cdc42显着促进SW480细胞通过小室迁移。结论我们的结果表明,活性Rho GTPase Cdc42可以大大增强结直肠癌细胞SW480的扩散,迁移和侵袭能力,这可能有助于结直肠癌的转移。

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