蟾毒灵抑制人前列腺癌PC3细胞迁移侵袭的机制研究  

Research on the mechanism of bufalin inhibiting migration and invasion of human prostate cancer PC3 cells

作　　者：王海临[1] 周雯敏 扶佳俐 叶阳娇 扈新[1] 高慧林[1] 张建业 WANG Hai-lin;ZHOU Wen-min;FU Jia-li;YE Yang-jiao;HU Xin;GAO Hui-lin;ZHANG Jian-ye(Department of Urology,Bayannur Hospital,Bayannur 015000,China;Guangdong Key Laboratory of Molecular Target and Clinical Pharmacology,School of Pharmacy,Guangzhou Medical University,Guangzhou 511436,China)

机构地区：[1]巴彦淖尔市医院泌尿外科,内蒙古自治区巴彦淖尔015000 [2]广州医科大学药学院,广东省分子靶标与临床药理学重点实验室,广东广州511436

出　　处：《药学学报》2022年第5期1361-1366,共6页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(U1903126);2020年国家级大学生创新创业训练计划(202010570032)。

摘　　要：本研究主要探讨了蟾毒灵(bufalin)对人前列腺癌PC3细胞的体外增殖、迁移和侵袭的抑制能力,初步探讨蟾毒灵抑制上皮-间充质转化(epithelial-mesenchymal transformation,EMT)过程的分子机制。采用MTT法评价PC3细胞活力;划痕和Transwell实验用于检测细胞的迁移和侵袭能力;采用Western blot实验检测EMT和整合素家族蛋白的表达情况。结果显示,蟾毒灵对人前列腺癌PC3细胞的半数抑制浓度值为0.26±0.03μmol·L-1,蟾毒灵处理后,PC3细胞的迁移速度变慢(P<0.05),穿过微孔滤膜的PC3细胞数量减少(P<0.05),这表明蟾毒灵能够浓度依赖性地抑制PC3细胞的增殖、迁移与侵袭能力。本研究发现蟾毒灵可以影响EMT相关蛋白的表达,包括E-钙黏附素(E-cadherin)的上调以及N-钙黏附素(N-cadherin)、β-连环蛋白(β-catenin)、基质金属蛋白酶9(MMP9)、基质金属蛋白酶2(MMP2)、癌基因c-myc、锌指转录因子Snail的下调;蟾毒灵还可以抑制整合素家族蛋白的表达,包括整合素α2(ITGA2)、整合素β1(ITGB1)、整合素β3(ITGB3)、整合素β5(ITGB5)、Yes相关蛋白/转录共激活因子PDZ结合基序(YAP/TAZ)和整合素连接激酶(ILK)。此外,蟾毒灵还可抑制磷酸化局部黏着斑激酶(p-FAK)/局部黏着斑激酶(FAK)、磷酸化类固醇受体辅激活因子(p-Src)/类固醇受体辅激活因子(Src)、磷酸化蛋白激酶B(p-Akt)/蛋白激酶B(Akt)的蛋白表达水平,上述研究结果表明,蟾毒灵可能通过FAK/Src/磷酸肌醇3-激酶(PI3K)/Akt通路抑制前列腺癌PC3细胞的增殖、转移和侵袭能力。因此,蟾毒灵为前列腺癌治疗药物的开发提供一定的参考价值。In this study,we investigated the inhibitory effects of bufalin on proliferation,migration and invasion of PC3 cells in vitro,and preliminarily explored the molecular mechanism of epithelial-mesenchymal transformation(EMT)inhibited by bufalin.The viability of PC3 cells was evaluated by MTT assay,and the migration and invasion abilities of PC3 cells were detected by wound healing and Transwell assay.Western blot was used to detect the expression of EMT and integrin family proteins.The results showed that the half maximal inhibitory concentration(IC50)value of bufalin against PC3 cells was 0.26±0.03μmol·L-1.After bufalin treatment,the migration rate of PC3 cells slowed down(P<0.05),the number of PC3 cells passing through the microporous membrane decreased(P<0.05),which indicated that bufalin could inhibit the proliferation,migration and invasion of PC3 cells in a concentration-dependent manner.We found that bufalin could affect the expression of EMT-related proteins,including up-regulation of E-cadherin and down-regulation of N-cadherin,β-catenin,matrix metalloproteinase 9(MMP9),matrix metalloproteinase 2(MMP2),c-myc and Snail.Bufalin also inhibited the expression of integrin family proteins,including integrinα2(ITGA2),integrinβ1(ITGB1),integrinβ3(ITGB3),integrinβ5(ITGB5),Yes-associated protein/transcriptional coactivator with a PDZ-binding motif(YAP/TAZ)and integrin-linked kinase(ILK).In addition,bufalin could also inhibit the protein expression level of phospho-focal adhesion kinase(p-FAK)/FAK,phospho-steroid receptor coactivator(p-Src)/Src and phospho-protein kinase B(p-Akt)/Akt.These results suggested that bufalin might inhibit the proliferation,metastasis and invasion of prostate cancer PC3 cells through the FAK/Src/phosphoinositide 3-kinase(PI3K)/Akt pathway.Therefore,bufalin provides reference value for the development of therapeutic drugs for prostate cancer.

关 键 词：蟾毒灵 前列腺癌 抗肿瘤 增殖 转移 

分 类 号：R966[医药卫生—药理学]