HC-1119对三阴性乳腺癌BT549细胞生物学行为的影响

Effect of HC-1119 on biological behaviors of triple-negative breast cancer BT549 cells

目的:检测新型人工合成的雄激素受体(androgen receptor,AR)拮抗剂HC-1119对三阴性乳腺癌BT549细胞生物学功能的影响,并探讨其分子机制。方法:用Western blot法检测不同人乳腺癌细胞系MDA-MB-231、T47D、MCF-7、SKBR3和BT549中AR的表达。在AR表达阳性的三阴性乳腺癌BT549细胞中,采用HC-1119进行刺激,用CCK-8法检测细胞活力;流式细胞术检测细胞凋亡和周期分布情况,Transwell实验检测细胞迁移和侵袭情况,Western blot法检测HC-1119刺激对BT549细胞中E-cadherin、vimentin和P21表达的影响。结果:AR在BT549细胞中阳性表达;HC-1119呈浓度和时间依赖性抑制BT549细胞的活力(P<0.05),显著上调BT549细胞的早期凋亡百分比和S期百分比(P<0.01),抑制BT549细胞的迁移和侵袭能力(P<0.05);此外,HC-1119还能抑制BT549细胞中P21表达(P<0.01),而E-cadherin和vimentin的表达未见改变。结论:AR拮抗剂HC-1119能够抑制AR阳性的三阴性乳腺癌BT549细胞的活力、迁移能力和侵袭能力,促进细胞凋亡,阻滞细胞周期;HC-1119通过P21调节细胞生长,却没有经过上皮-间充质转化而参与BT549细胞的迁移行为。

AIM:To explore the effect of new artificially synthesized androgen receptor(AR)antagonist HC-1119 on the biological function of triple-negative breast cancer(TNBC)BT549 cells and the molecular mechanism.METHODS:The AR expression was assessed in different human breast cancer cell lines MDA-MB-231,T47D,MCF-7,SKBR3 and BT549 by Western blot.The TNBC BT549 cells with AR positive expression were treated with HC-1119.The cell viability was measured by CCK-8 assay.The apoptosis rate and cell cycle distribution were analyzed by flow cytometry.The migration and invasion abilities were detected by Transwell assay in vitro.The protein expression of E-cadherin,vimentin and P21 was determined by Western blot.RESULTS:AR was positively expressed in BT549 cells.HC-1119 inhibited the cell viability in a time-and dose-dependent manner(P<0.05),increased the percentage of apoptotic cells and the percentage of S-phase cells significantly,repressed the migration and invasion abilities(P<0.05),and decreased P21 expression at protein level(P<0.01).No influence on the expression of E-cadherin and vimentin in the BT549 cells was observed.CONCLUSION:AR antagonist HC-1119 decreases the viability,migration ability and invasion ability,enhances the apoptosis,and arrests the cell cycle distribution of TNBC BT549 cells.HC-1119 represses the viability of BT549 cells by down-regulating P21 expression,while the process of epithelial-mesenchymal transition is not involved in the inhibition of cell migration.