Cynanoside H对三阴性乳腺癌MDA-MB-231细胞增殖和转移的影响及其机制  

Effect of Cynanoside H on proliferation and metastasis of triple negative breast cancer MDA⁃MB⁃231 cells and its mechanism

作　　者：赵鹏 李艳梅 邱剑飞 潘朝兰 杨珏 郝小江 ZHAO Peng;LI Yanmei;QIU Jianfei;PAN Chaolan;YANG Jue;HAO Xiaojiang(School of Pharmaceutical Sciences,Guizhou Medical University,Anshun 561113,Guizhou Province,China;不详)

机构地区：[1]贵州医科大学药学院,贵州安顺561113 [2]贵州医科大学省部共建药用植物功效与利用国家重点实验室,贵州贵阳550014 [3]贵州省天然产物研究中心,贵州贵阳550014 [4]贵州医科大学基础医学院现代病原生物学特色重点实验室,贵州安顺561113

出　　处：《中国生物制品学杂志》2024年第10期1190-1199,共10页Chinese Journal of Biologicals

基　　金：国家自然科学基金地区项目(NSFC:82160813);贵州省省级科技计划项目资助(黔科合基础-ZK[2021]一般526)。

摘　　要：目的探讨Cynanoside H对三阴性乳腺癌(triple negative breast cancer,TNBC)MDA-MB-231细胞增殖和转移的影响及其作用机制,为治疗TNBC药物的开发提供实验依据。方法将MDA-MB-231细胞随机分为对照组(不加Cynanoside H)和不同浓度Cynanoside H(2.5、5、10μmol/L)处理组。采用MTT法、平板克隆形成法检测Cynanoside H对MDA-MB-231细胞增殖的影响;流式细胞术检测Cynanoside H对MDA-MB-231细胞周期的影响;划痕-愈伤试验、Transwell迁移及侵袭试验检测Cynanoside H对MDA-MB-231细胞转移的影响;Western blot检测Cynanoside H对MDAMB-231细胞中周期(c-Myc、CDK1、CDK2及cyclin E1)、转移(E-cadherin、Vimentin及β-catenin)及PDGFRB/JAK2/STAT3通路相关蛋白(PDGFRB、p-JAK2、JAK2、p-STAT3及STAT3)表达的影响。结果2.5、5、10μmol/L Cynanoside H剂量组的细胞活力(t分别为4.598、19.77和53.43,P均<0.05)、细胞生长(36 h:t分别为8.256、11.57和12.16,P均<0.05;48 h:t分别为10.49、22.49和19.63,P均<0.01;72 h:t分别为50.20、28.84和15.83,P均<0.01;96 h:t分别为11.18、18.35和20.92,P均<0.01)及细胞克隆形成(t分别为4.618、9.821和12.14,P均<0.05)均明显低于对照组。与对照组相比,Cynanoside H各剂量组以浓度依赖性方式增加了S期的细胞比例(48 h:t分别为6.316、8.156和14.11,P均<0.05;72 h:t分别为7.231、15.36和25.16,P均<0.05),且下调了c-Myc(5和10μmol/L:t分别为10.39和12.18,P<0.05和<0.01)、CDK1(t分别为3.777、5.069和6.974,P均<0.05)、CDK2(5和10μmol/L:t分别为12.72和19.43,P均<0.01)及cyclin E1(5和10μmol/L:t分别为3.813和15.23,P均<0.01)在蛋白水平的表达。Cynanoside H各剂量组与对照组相比,明显抑制了MDA-MB-231细胞划痕愈合(48 h:t分别为6.969、56.16和27.73,P均<0.05;72 h:t分别为8.619、22.12和32.15,P均<0.05)、Trswell迁移(t分别为9.817、14.74和19.39,P均<0.01)及侵袭(t分别为5.614、13.85和14.22,P均<0.01),且上调了E-cadherin在蛋白水平的表达(10μmol/L:t=11.79,P<0.01),下调了VimentiObjective To investigate the effect of Cynanoside H on the proliferation and metastasis of triple negative breast cancer(TNBC)MDA-MB-231 cells and its mechanism,so as to provide an experimental basis for the development of TNBC therapeutics.Methods MDA-MB-231 cells were randomly divided into one control group(without Cynanoside H)and three test groups including 2.5,5 and 10μmol/L Cynanoside H dose groups.The effect of Cynanoside H on the proliferation of MDA-MB-231 cells was detected by MTT assay and clone formation assay,while the effect on the cell cycle was detected by flow cytometry,and the effect on the metastasis of MDA-MB-231 cells was measured by wound healing assay,Transwell migration and invasion assay.In addition,the effects of Cynanoside H on the expression of cell cycle(c-Myc,CDK1,CDK2 and cyclin E1),metastasis(E-cadherin,Vimentin andβ-catenin)and PDGFRB/JAK2/STAT3 pathway related proteins(PDGFRB,p-JAK2,JAK2,p-STAT3 and STAT3)in MDA-MB-231 cells were determined by Western blot.Results The levels of cell viability(t=4.598,19.77 and 53.43,respectively,each P<0.05),growth(36 h:t=8.256,11.57 and 12.16,respectively,each P<0.05;48 h:t=10.49,22.49 and 19.63,respectively,each P<0.01;72 h:t=50.20,28.84 and 15.83,respectively,each P<0.01;96 h:t=11.18,18.35 and 20.92,respectively,each P<0.01)and clone formation(t=4.618,9.821 and 12.14,respectively,each P<0.05)of 2.5,5 and 10μmol/L dose groups were significantly lower than those of the control group.Compared with the control group,the proportion of cells in S phase of three test group increased in a concentration-dependent manner(48 h:t=6.316,8.156 and 14.11,respectively,each P<0.05;72 h:t=7.231,15.36 and 25.16,respectively,each P<0.05),and the expression of c-Myc(5 and 10μmol/L:t=10.39 and 12.18,P<0.05 and<0.01,respectively),CDK1(t=3.777,5.069 and 6.974,respectively,each P<0.05),CDK2(5 and 10μmol/L:t=12.72 and 19.43,respectively,each P<0.01)and cyclinE1(t=3.813 and 15.23,respectively,each P<0.01)was down-regulated at the protein level.Compared with the con

关 键 词：Cynanoside H 三阴性乳腺癌 MDA-MB-231细胞 细胞增殖 细胞转移 

分 类 号：R962[医药卫生—药理学]