氯氧喹通过下调Rho/Rho激酶信号通路抑制乳腺癌细胞转移

目的: 通过观察氯氧喹对乳腺癌细胞系细胞骨架的聚合状态及其对Rho/Rho激酶信号通路相关靶点蛋白磷酸化的作用，明确氯氧喹治疗乳腺癌的作用机制。方法: 以不同浓度氯氧喹处理乳腺癌Bcap37和MDA-MB-453细胞系，划痕试验检测细胞迁移，Transwell试验检测细胞侵袭，罗丹明标记鬼笔环肽染色法观察F肌动蛋白的聚解状态，免疫荧光法观察α微管蛋白的表达，蛋白质印迹法检测Rho/Rho激酶信号通路关键蛋白磷酸化水平。结果: 氯氧喹可剂量依赖性地抑制细胞迁移和侵袭；Bcap37和MDA-MB-453细胞经氯氧喹处理后其细胞形态发生改变，细胞体积减少；F肌动蛋白和α微管蛋白染色形态不规则，荧光染色聚集成团，且呈现剂量依赖性；氯氧喹可下调Rho/Rho激酶信号通路中肌动蛋白解聚因子Cofilin、磷酸化蛋白激酶（Limk）、Rho关联卷曲蛋白激酶2（Rock2）磷酸化表达（均P < 0.01）。结论: 氯氧喹可抑制乳腺癌细胞骨架聚合从而抑制细胞迁移，该作用可能与抑制Rho/Rho激酶信号通路相关。

Chloroxoquinoline inhibits invasion in breast cancer via down-regulating Rho/Rho kinase signaling pathway

Objective: To investigate the effect of chloroxoquinoline on cytoskeleton of breast cancer cells and its relation with Rho/Rho kinase signaling pathway. Methods: Breast cancer Bcap37 and MDA-MB-453 cells were treated with different concentrations of chloroxoquinoline. Wound healing and Transwell assay were conducted to detect cell migration and invasion, respectively. Rhodamine-phalloidin staining and immunofluorescent staining were used to observe the polymerization state of F-actin and the expression of α-Tublin in breast cancer cells, respectively. Western blot was used to detect the phosphorylation level of key protein in Rho/Rho kinase signaling pathway. Results: Compared with the control group, chloroxoquinoline treatment induced dose-dependent decrease in cell migration and invasion, and Bcap37 and MDA-MB-453 cells treated with chloroxoquinoline showed dose-dependent changes in cell morphology and decrease in cell body. The staining of F-actin and α-Tublin was irregular and clustered. Furthermore, treatment of chloroxoquinoline down-regulated the phosphorylation of the Rho/Rho kinase signaling proteins Cofilin, Limk and Rock2 (all P < 0.01). Conclusions: Chloroxoquinoline inhibits the cytoskeleton in breast cancer Bcap37 and MDA-MB-453 cells and inhibits cell migration. This effect may be associated with down-regulation of Rho/Rho kinase signaling pathway.