摘要
目的探讨心房利钠肽(atrial natriuretic peptide,ANP)对人三阴性乳腺癌细胞株MDA-MB-231增殖和周期的影响。方法在MDA-MB-231细胞中加入不同浓度ANP,分为对照组(ANP 0μmol·L^-1)和实验组(ANP 1μmol·L^-1、ANP 10μmol·L^-1、ANP 100μmol·L^-1),MTT法检测ANP对细胞抑制作用,细胞克隆实验检测细胞集落形成能力,流式细胞仪(FCM)检测细胞周期,蛋白免疫印迹法(Western blot)检测CyclinB1、CyclinD1、CDK1、CDK4蛋白相对表达量。结果MTT法结果表明,实验组细胞的生长抑制率均较对照组增加(P均<0.01)。克隆实验显示,随着ANP浓度增加,集落形成数逐渐降低(P<0.05)。FCM检测提示,与对照组相比,实验组细胞在G0/G1期增加,S期和G2/M期占比有不同程度下降(P均<0.05)。Western blot结果显示,实验组较对照组CyclinB1、CyclinD1、CDK1、CDK4蛋白表达水平下调(P均<0.05)。结论ANP可抑制MDA-MB-231增殖,诱导细胞阻滞于G0/G1期,通过降低CyclinB1、CyclinD1、CDK1、CDK4蛋白表达调控细胞周期。
Objective To investigate the effect of atrial natriuretic peptide(ANP)on the proliferation and cycle of human three negative breast cancer cell line MDA-MB-231.Methods Different concentrations of ANP were added into the MDA-MB-231 cells,and the experiment was divided into control group(ANP 0μmol·L^-1)and experimental group(ANP 1μmol·L^-1,ANP10μmol·L^-1,ANP100μmol·L^-1),and the inhibition of cell apoptosis was detected by MTT,The colony forming ability was detected by cell clone experiment.Flow cytometry(FCM)was used to detect cell cycle,and Western Blot was used to detect the relative expression of CyclinB1,CyclinD1,CDK1 and CDK4.Results MTT method showed that inhibitory effect on cells increased in the experimental group than in the control group(P<0.01).Colony formation number gradually decreased(P<0.05).FCM test showed that compared with the control group,the cells in the experimental group increased significantly in the G0/G1 phase,and the S phase and G2/M phase decreased to varying degrees(P all<0.05)showed a concentration correlation.Western blot showed that the expression level of CyclinB1,CyclinD1,CDK1 and CDK4 protein decreased in the experimental group than in the control group(P all<0.05).Conclusion ANP can inhibit MDA-MB-231 proliferation,induce cell arrest in G0/G1 phase,and regulate cell cycle by decreasing CyclinB1,CyclinD1,CDK1 and CDK4 protein expression.
作者
张弛
陆俊铭
周炳刚
曹相玫
ZHANG Chi;LU Junming;ZHOU Binggang;CAO Xiangmei(The Second Affiliated Hospital of Ningxia Medical University,Yinchuan 750001,China;Department of Pathology,School of Basic Medical Sciences,Ningxia Medical University,Yinchuan 750004,China)
出处
《宁夏医科大学学报》
2020年第5期452-456,共5页
Journal of Ningxia Medical University
基金
国家自然科学基金(81560501)
宁夏自然科学基金(NZ17238)。
