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J Zhejiang Univ (Med Sci)  2019, Vol. 48 Issue (6): 631-637    DOI: 10.3785/j.issn.1008-9292.2019.12.07
Chloroxoquinoline inhibits invasion in breast cancer via down-regulating Rho/Rho kinase signaling pathway
LIU Jingwen1(),YANG Xinglian1,SHEN Kaili1,ZENG Linghui1,*(),SUN Yan2,*()
1. School of Medicine, Zhejiang University City College, Hangzhou 310015, China
2. Department of Oncology, Zhejiang Cancer Hospital, Hangzhou 310022, China
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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.

Key wordsBreast neoplasms/pathology      Cytoskeleton      rho-Associated kinases/antagonists & inhibitors      Signal transduction/drug effects      Actins      Tubulin      Antineoplastic agents/pharmacology     
Received: 15 July 2019      Published: 19 January 2020
CLC:  R737.9  
Corresponding Authors: ZENG Linghui,SUN Yan     E-mail:;;
Cite this article:

LIU Jingwen,YANG Xinglian,SHEN Kaili,ZENG Linghui,SUN Yan. Chloroxoquinoline inhibits invasion in breast cancer via down-regulating Rho/Rho kinase signaling pathway. J Zhejiang Univ (Med Sci), 2019, 48(6): 631-637.

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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。

关键词: 乳腺肿瘤/病理学,  细胞骨架,  rho相关激酶类/拮抗剂和抑制剂,  信号传导/药物作用,  肌动蛋白类,  微管蛋白,  抗肿瘤药/药理学 
Fig 1 The results of cell scratch test in MDA-MB- 453 cells
Fig 2 The results of Transwell test in MDA-MB- 453 cells
Fig 3 Distribution of cytoskeletal actin in Bcap37 and MDA-MB- 453 cells (Phalloidin-iFluor 488)
Fig 4 Distribution of α-microtubulin in Bcap37 and MDA-MB- 453 cells (immunofluorescence staining)
Fig 5 Representative blots of protein expression of Rho/Rho kinase signaling in MDA-MB- 453 cells
Fig 6 Statistic analysis of phosphorylated protein expression of Rho/Rho kinase signaling protein in MDA-MB- 453 cells (n=6)
[1]   YOULDEN D R , CRAMB S M , YIP C H et al. Incidence and mortality of female breast cancer in the Asia-Pacific region[J]. Cancer Biol Med, 2014, 11 (2): 101- 115
[2]   SUNG H , ROSENBERG P S , CHEN W Q et al. Female breast cancer incidence among Asian and Western populations:more similar than expected[J]. J Natl Cancer Inst, 2015, 107 (7): pii:djv107
[3]   耿维凤 . 抗癌新药-氯氧喹胶囊[J]. 中国医院药学杂志, 2005, 25 (10): 994
GENG Weifeng . Anticancer drug-chloroxoquinoline capsule[J]. Chinese Journal of Hospital Pharmacy, 2005, 25 (10): 994
doi: 10.3321/j.issn:1001-5213.2005.10.055
[4]   孙燕, 冯琳迤, 雷蕾 et al. 氯氧喹对不同类型乳腺癌细胞系的抑制作用及机制[J]. 中国药理学通报, 2019, 35 (3): 347- 353
SUN Yan , FENG Linyi , LEI Lei et al. Inhibitory effect of chloroxoquinoline in different breast cancer cells[J]. Chinese Pharmacological Bulletin, 2019, 35 (3): 347- 353
doi: 10.3969/j.issn.1001-1978.2019.03.011
[5]   OLSON M F . Applications for ROCK kinase inhibition[J]. Current Opinion in Cell Biology, 2008, 20 (2): 242- 248
doi: 10.1016/
[6]   肖英, 程爱兰 . 细胞骨架在肿瘤侵袭转移中的研究进展[J]. 中国肿瘤临床, 2016, 43 (22): 1007- 1011
XIAO Ying , CHENG Ailan . Cell cytoskeleton-the movers in cancer metastasis[J]. Chinese Journal of Clinical Oncology, 2016, 43 (22): 1007- 1011
doi: 10.3969/j.issn.1000-8179.2016.22.828
[7]   ROTTNER K , STRADAL T E . Actin dynamics and turnover in cell motility[J]. Curr Opin Cell Biol, 2011, 23 (5): 569- 578
doi: 10.1016/
[8]   AYLETT C H , L?WE J , AMOS L A . New insights into the mechanisms of cytomotive actin and tubulin filaments[J]. Int Rev Cell Mol Biol, 2011, 292 1- 71
doi: 10.1016/B978-0-12-386033-0.00001-3
[9]   FRANCIS R, XU X, PARK H, et al. Connexin43 modulates cell polarity and directional cell migration by regulating microtubule dynamics[J/OL]. PLoS One, 2011, 6(10): e26379.
[10]   KATSETOS C D , DRáBEROVá E , LEGIDO A et al. Tubulin targets in the pathobiology and therapy of glioblastoma multiforme. Ⅱ. γ-Tubulin[J]. J Cell Physiol, 2009, 221 (3): 514- 520
doi: 10.1002/jcp.21884
[11]   PRICE L S , COLLARD J G . Regulation of the cytoskeleton by Rho-familyGTPases:implications for tumour cell invasion[J]. Semin Cancer Biol, 2001, 11 (2): 167- 173
doi: 10.1006/scbi.2000.0367
[12]   TAKAHASHI N , NOBUSUE H , SHIMIZU T et al. ROCK inhibition induces terminal adipocyte differentiation and suppresses tumorigenesis in chemoresistant osteosarcoma cells[J]. Cancer Res, 2019, 79 (12): 3088- 3099
doi: 10.1158/0008-5472.CAN-18-2693
[13]   CASCIONE M , DE MATTEIS V , TOMA C C et al. Morphomechanical and structural changes induced by ROCK inhibitor in breast cancer cells[J]. Exp Cell Res, 2017, 360 (2): 303- 309
doi: 10.1016/j.yexcr.2017.09.020
[14]   HUANG G X , WANG Y , SU J et al. Up-regulation of Rho-associated kinase 1/2 by glucocorticoids promotes migration, invasion and metastasis of melanoma[J]. Cancer Lett, 2017, 410 1- 11
doi: 10.1016/j.canlet.2017.09.005
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