肌动蛋白调节分子CAP1对肺癌细胞迁移和增殖的影响

Adenylatecyclase-associated protein 1 enhances motility and promotes proliferation of non-small cell lung cancer cells in vitro

目的探讨腺苷酸环化酶关联蛋白1(adenylatecyclase-associated protein 1,CAP1)在非小细胞肺癌(non-small cell lung cancer,NSCLC)中的表达及其对肺癌细胞迁移和增殖的影响。方法收集本院2010-2015年NSCLC临床标本42例,采用免疫组化检测肺癌组织、癌旁组织和正常肺组织中CAP1的表达;以sh RNA慢病毒感染NSCLC细胞系,敲低CAP1表达,采用划痕和Transwell实验观测敲低CAP1后肺癌细胞迁移能力的变化;MTS法及克隆形成实验检测细胞增殖能力的变化;免疫荧光观察细胞骨架的变化;流式细胞仪检测细胞周期的改变。结果与癌旁组织和正常肺组织相比,CAP1蛋白在NSCLC癌组织中显著高表达(P<0.01);划痕实验及Transwell实验显示,敲低CAP1的肺癌细胞较对照细胞迁移能力显著减弱(P<0.01),细胞肌动蛋白骨架改变,并且其增殖减慢,克隆形成能力降低(P<0.01);细胞周期分析发现CAP1敲低的肺癌细胞阻滞在G0/G1期。结论 CAP1促进肺癌细胞的迁移和增殖,可能与CAP1参与细胞肌动蛋白骨架和细胞周期调控有关。

Objective To investigate the expression of adenylatecyclase-associated protein 1（ CAP1）in human non-small cell lung cancer（ NSCLC） and its role in the migration and proliferation of NSCLC cells.Methods Immunohistochemistry was used to detect the expression of CAP1 protein in clinical specimens of NSCLC,adjacent tissues and normal lung tissues from 42 patients. Two human NSCLC cell lines（ A549 and H460） with CAP1 knockdown by sh RNA lentivirus infection were examined for changes in cell motility using scratch wound healing test and Transwell assay,and the cell proliferation was assessed using MTS test and colony formation assay; and changes of cytoskeleton were observed with immunofluorescence assay and the cell cycle changes were analyzed using flow cytometry. Results The clinical specimens of NSCLC showed obvious CAP1 overexpression as compared with the adjacent tissues and normal lung tissues（ P〈0. 01）.Knockdown of CAP1 significantly impaired the migration（ P〈0. 01） and proliferation of NSCLC cells in vitro,resulting also in depressed clone formation ability（ P〈0. 01）,obvious changes in actin cytoskeleton,and cell cycle arrest in G0/G1 phase. Conclusion CAP1 promotes the motility and proliferation of NSCLC cells possibly by regulating actin cytoskeleton and cell cycle.