桃叶珊瑚苷对乳腺癌细胞恶性生物学行为的影响及机制

Effects and mechanism of aucubin on malignant biological behavior of breast cancer cells

目的探讨桃叶珊瑚苷(Auc)调节周期蛋白依赖性激酶1(CDK1)/周期蛋白B1(CCNB1)/Polo样激酶1(PLK1)信号通路对乳腺癌细胞恶性生物学行为的影响。方法以人乳腺癌细胞MCF-7为对象,将其分为对照组,Auc低、中、高浓度组(Auc-L、Auc-M、Auc-H组,20、40、80μmol/L的Auc),Auc-H+pcDNA-NC组(80μmol/L的Auc+转染pcDNA-NC质粒),Auc-H+pcDNA-CDK1组(80μmol/L的Auc+转染pcDNA-CDK1质粒),检测各组细胞的增殖、克隆形成、侵袭、迁移能力,凋亡情况及周期分布,凋亡、上皮-间充质转化(EMT)、CDK1/CCNB1/PLK1信号通路相关蛋白的表达情况。以BALB/c裸鼠为对象,通过皮下接种MCF-7细胞悬液建立移植瘤模型,并将其分为对照组和Auc组(每组12只),检测各组裸鼠肿瘤体积、质量及肿瘤组织中CDK1/CCNB1/PLK1信号通路相关蛋白的表达情况。结果与对照组细胞相比,Auc各浓度组的细胞克隆形成数、增殖率、细胞侵袭数、划痕愈合率、G1/G0期和S期细胞占比以及B细胞淋巴瘤2(Bcl-2)、神经钙黏素、纤维连接蛋白、CDK1、CCNB1、PLK1蛋白的表达水平均显著降低(P<0.05),细胞凋亡率、G2/M期细胞占比以及Bcl-2相关X蛋白、上皮钙黏素蛋白的表达水平均显著增加(P<0.05),呈浓度依赖性(P<0.05);与Auc-H+pcDNA-NC组细胞相比,Auc-H组细胞上述指标的差异均无统计学意义(P>0.05),而Auc-H+pcDNA-CDK1组细胞上述指标则显著逆转(P<0.05)。与对照组裸鼠相比,Auc组裸鼠肿瘤体积、质量以及肿瘤组织中CDK1、CCNB1、PLK1蛋白的表达水平均显著降低(P<0.05)。结论Auc能抑制乳腺癌细胞的增殖、侵袭和迁移,诱导细胞周期阻滞,抑制EMT进程,该作用可能与抑制CDK1/CCNB1/PLK1信号通路激活有关。

OBJECTIVE To investigate the effects of aucubin(Auc)on the malignant biological behavior of breast cancer cells by regulating cyclin-dependent kinase 1(CDK1)/cyclin B1(CCNB1)/Polo-like kinase 1(PLK1)signaling pathway.METHODS Human breast cancer cells MCF-7 were divided into control group,Auc low-,medium-and high-concentration groups(AUC-L,AUC-M,AUC-H groups,20,40 and 80μmol/L Auc),Auc-H+pcDNA-NC group(80μmol/L Auc+transfected pcDNA-NC plasmid),and Auc-H+pcDNA-CDK1 group(80μmol/L Auc+transfected pcDNA-CDK1 plasmid).Cell proliferation,clonal formation,invasion and migration abilities,apoptosis and cycle distribution,and the expressions of related proteins of apoptosis,epithelial-mesenchymal transformation(EMT)and CDK1/CCNB1/PLK1 signaling pathway were detected in each group.The transplanted tumor model of BALB/c nude mice was established by subcutaneous inoculation of MCF-7 cell suspension,and the mice were divided into control group and Auc group(12 mice in each group).The tumor volume,mass and the expressions of related proteins of CDK1/CCNB1/PLK1 signaling pathway in tumor tissues were detected.RESULTS Compared with control group,the number of clonal formation,proliferation rate,cell invasion number,scar healing rate,G1/G0 phase and S phase cell proportions,and the expressions of B cell lymphoma-2(Bcl-2),N-cadherin,fibronectin,CDK1,CCNB1 and PLK1 were decreased significantly(P<0.05).The apoptotic rate,G2/M phase cell proportion and the expressions of Bcl-2 associated X protein and E-cadherin were significantly increased,in a dose-dependent manner(P<0.05).Compared with the Auc-H+pcDNA-NC group,there was no statistical significance in the above indexes in the Auc-H group(P>0.05),while the above indexes in the Auc-H+pcDNA-CDK1 group were significantly reversed(P<0.05).Compared with the control group,the tumor volume and mass,and the expressions of CDK1,CCNB1 and PLK1 in tumor tissue of Auc group were significantly decreased(P<0.05).CONCLUSIONS Auc can inhibit the proliferation,invasion and migration of breast cancer cells,induce cell cycle arrest,and inhibit the progression of EMT,which may be related to inhibiting the activation of the CDK1/CCNB1/PLK1 signaling pathway.