氯氧喹对不同类型乳腺癌细胞系的抑制作用及机制

Inhibitory effect of chloroxoquinoline in different breast cancer cells

目的探究氯氧喹对不同类型乳腺癌细胞增殖、凋亡及细胞周期阻滞的作用及其可能机制。方法采用不同浓度氯氧喹处理乳腺癌Bcap37、MDA-MB-231和MDA-MB-453细胞。MTT法检测细胞活力,Annexin V-/PI双染法采用流式细胞仪检测细胞凋亡和细胞周期,蛋白印迹法检测凋亡和细胞周期相关蛋白水平的变化。结果氯氧喹在50 mg·L^(-1)以上对3种细胞系的生长均有明显抑制作用(P<0.05),并呈剂量依赖性,且200 mg·L^(-1)氯氧喹与40 mg·L^(-1)紫杉醇作用相近。氯氧喹可明显诱导3种细胞凋亡(P<0.05),且对MDA-MB-231细胞凋亡的作用尤其明显(P<0.01),并与凋亡相关蛋白cleaved-PARP、cleaved-caspase的表达相一致。氯氧喹还可明显诱导3种细胞G_2/M期的数目增加(P<0.05),但在Bcap37和MDA-MB-453细胞更明显,并伴随p21蛋白下调和CDC2蛋白的上调。结论氯氧喹能有效抑制乳腺癌细胞的增殖活力,在MDA-MB-231细胞以促进凋亡为主,而在Bcap37和MDA-MB-453细胞以细胞周期阻滞为主。

Aim To explore the effect of chloroxoquinoline(Chl)on cell proliferation,apoptosis and cell cycle in breast cancer cells and its potential mechanism.Methods Breast cancer cells of Bcap37,MDA-MB-231and MDA-MB-453 were treated with Chl at different doses.Cell proliferation was detected by MTT assay.Cell apoptosis and cell cycle were assayed by Annexin V-/PI kit using flow cytometry.Protein expression related to apoptosis and cell cycle were determined by Western blot.Results Chl over the dose of 50 mg·L^-1 inhibited cell proliferation in all three cell lines in a dose-dependent manner,and Chl at the dose of 200 mg·L^-1 exhibited similar efficiency as 40 mg·L^-1 Paclitaxel did.When the cells were treated with 100 or 200 mg·L^-1 Chl for 48 h,the apoptotic ratio was dose-dependently up-regulated in all three cell lines(P<0.05),especially in the MDA-MB-231cells.It was further proved by the protein expression of cleaved-PARP and cleaved-caspase related to apoptosis.On the contrary,Chl at the dose of 100mg·L^-1 significantly increased the cell number of G2/M stage in Bcap37 and MDA-MB-453 cells(P<0.05).Correspondingly,Western blot analysis demonstrated that Chl dose-dependently down-regulated p21 protein and up-regulated CDC2 protein,especially in Bcap37 and MDA-MB-453 cells.Conclusions Chl significantly inhibits cell proliferation in breast cancer cells of Bcap37,MDA-MB-231 and MDA-MB-453,which may mainly result from its induction of cell apoptosis in MDA-MB-231 cells and cell cycle arrest in Bcap37 and MDA-MB-453 cells.These data provide new idea of chemotherapy using Chl for breast cancer treatment.