小分子化合物Zinc-09通过诱导MDA-MB-453细胞G1期阻滞抗乳腺癌作用机制研究  

Effect and mechanism of small molecular compound Zinc-09 on anti breast cancer via induce G1 phase arrest of MDA-MB-453 cells

作　　者：于晓萍 周端方 周维英 王雅蘭 李虹瑶 李慧珍 曾鸿芳 宋燚 李小丽 YU Xiao-ping;ZHOU Duan-fang;ZHOU Wei-ying;WANG Ya-lan;LI Hong-yao;LI Hui-zhen;ZENG Hong-fang;SONG Yi;LI Xiao-li(Department of Pharmacology,College of Pharmacy,Chongqing Medical University,Chongqing 400016,China;Chongqing Key Laboratory of Drug Metabolism,Chongqing Medical University,Chongqing 400016,China;Key Laboratory for Biochemistryand Molecular Pharmacology of Chongqing,Chongqing Medical University,Chongqing 400016,China;College of Pharmacy,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学,药学院药理学教研室,重庆400016 [2]重庆医科大学,重庆市高校生物化学与分子药理学重点实验室,重庆400016 [3]重庆医科大学,重庆市药物代谢研究重点实验室,重庆400016 [4]重庆医科大学,药学院,重庆400016

出　　处：《中国临床药理学杂志》2021年第22期3083-3087,共5页The Chinese Journal of Clinical Pharmacology

基　　金：重庆市教育委员会科学技术研究基金资助项目(KJQN201800431);重庆市自然科学基金面上基金资助项目(cstc2020jcyj-msxmX0223);重庆市高校创新研究群体基金资助项目(CXQT20012);重庆市研究生导师团队建设基金资助项目(dstd201807)。

摘　　要：目的研究小分子化合物Zinc-09(6-[2-(3,4-二甲氧基苯基)乙烯基]-5-硝基-2,4(1H,3H)-嘧啶二酮)体内外抗乳腺癌细胞MDA-MB-453的作用及机制。方法(1)细胞实验:将乳腺癌MDA-MB-453细胞分为4组:对照组(0.1%DMSO)和低、中、高3个浓度实验组(Zinc-09:50,100,200μmol·L^(-1)),处理细胞72 h,用四甲基偶氮唑蓝法检测细胞活力,流式细胞术检测细胞周期G1期的占比,蛋白质印迹法检测细胞周期蛋白依赖性激酶4(CDK4)的表达水平。(2)动物实验:体内建立裸鼠原位移植瘤模型,按肿瘤体积随机分成2组:模型组(2.0%DMSO)和Zinc-09组(50 mg·kg^(-1)),每组5只。连续给药21 d,记录并计算瘤体积和体重。结果(1)对照组和低、中、高3个浓度实验组的细胞活力分别为(100.00±1.00)%,(63.64±3.46)%,(38.78±5.18)%和(14.04±2.28)%;这4组处于G1期的比例分别为(58.58±1.29)%,(69.29±0.12)%,(72.33±0.45)%和(73.21±2.07)%;这4组细胞CDK4蛋白的相对表达量分别为1.05±0.02,0.77±0.02,0.43±0.01和0.28±0.01。上述指标:给药组与对照组比较,差异均有统计学意义(P<0.05,P<0.01)。(2)模型组和Zinc-09组的裸鼠肿瘤体积分别为(152.03±23.44)和(92.48±14.26)mm^(3),在9至21 d的5个时间点,2组间比较肿瘤体积差异均有统计学意义(P<0.05,P<0.01)。结论Zinc-09具有较好的体内外抗乳腺癌MDA-MB-453细胞的作用,其作用机制可能是通过诱导细胞G1期阻滞,从而发挥抗乳腺癌MDA-MB-453细胞的作用。Objective To study the effect and mechanism of small molecular compound Zinc-09(6-[2-(3.4-dimethoxyphenyl)vinyll-5-nitro-2.4(1H.3H)-pyrimidin-edione)on anti breast cancer MDA-MB-453 cell line in vivo and in vitro.Methods(1)Cell experiments:The breast cancer MDA-MB-453 cells were divided into 4 groups:control group(0.1%DMSO)and low,medium and high concentration experimental groups(50,100,200μmol·L^(-1) Zinc-09).After treating the cells for 72 h,the cell viability was measured by the tetramethylazolyl blue method;the proportion of the G1 phase of the cell cycle was measured by flow cytometry;the expression level of Cyclin-dependent kinase 4(CDK4)protein was measured by Western blot.(2)A nimal experiments:The orthotopic xenograft tumor of nude mouse was established in vivo,and mice were randomly divided into 2 groups according to the tumor volume:model group(2.0%DMSO)and Zinc-09 group(50 mg·kg^(-1)),5 mice/group.Each group was administered continuously for 21 d,then tumor volume and weight were recorded and calculated.Results(1)The cell viability of the control group and low,medium and high concentration experimental groups were(100.00±1.00)%,(63.64±3.46)%,(38.78±5.18)%and(14.04±2.28)%,respectively;the rates of G1 phase in the four groups were(58.58±1.29)%,(69.29±0.12)%,(72.33±0.45)%and(73.21±2.07)%;the relative expression levels of CDK4 protein in the four groups of cells were 1.05±0.02,0.77±0.02,0.43±0.01 and 0.28±0.01,respectively.Comparison between experimental groups and control group,the difference of indicators were significant(P<0.05,P<0.01).(2)The tumor volume of the nude mice in the model group and Zinc-09 group were(152.03±23.44)and(92.48±14.26)mm^(3),and the difference of tumor volume between the two groups at 5 time points from 9 to 21 d were significant(P<0.05,P<0.01).Conclusion Zinc-09 has anti-tumor effect on breast cancer in vitro and in vivo,and the mechanism of Zinc-09 may be relate to G1 phase arrest.

关 键 词：乳腺癌 细胞周期蛋白依赖性激酶4 细胞周期蛋白D1 细胞周期蛋白依赖性激酶抑制蛋白1A 6-[2-(3 4-二甲氧基苯基)乙烯基]-5-硝基-2 4(1H 3H)-嘧啶二酮(Zinc-09) 

分 类 号：R97[医药卫生—药品]