摘要
目的探讨秦皮甲素对三阴性乳腺癌细胞增殖的影响及其分子机制。方法分别以28、56、112、225、450和900μmol/L的秦皮甲素处理三阴性乳腺癌MDA-MB-231细胞24、48和72 h,同时设空白对照组,采用细胞计数试剂盒8(CCK-8)法检测细胞活力。分别以225、450和900μmol/L的秦皮甲素处理MDA-MB-231细胞48 h,同时设空白对照组,采用倒置显微镜观察细胞的形态变化,采用克隆形成实验检测克隆形成能力,采用实时荧光定量聚合酶链反应检测细胞中人类免疫缺陷病毒短转录诱导物连接因子1(FBI-1)、p53和p21 mRNA的表达水平,采用Western blot法检测细胞中FBI-1、p53、p21和Ki67蛋白的表达水平。结果与空白对照组相比,经28、56、112、225、450和900μmol/L秦皮甲素处理后MDA-MB-231细胞活性均明显降低,并呈浓度依赖性和时间依赖性。225、450和900μmol/L秦皮甲素处理后,MDA-MB-231细胞皱缩变扁,贴壁差,并出现不完整细胞,脱落,细胞间距变大,其中900μmol/L秦皮甲素组细胞变化最明显;与空白对照组比较,225、450和900μmol/L秦皮甲素组细胞克隆形成率均降低[分别为(77.18±5.13)%、(65.94±4.98)%和(45.92±3.70)%,均P<0.01],FBI-1 mRNA及蛋白表达增高(均P<0.01),p53和p21 mRNA及蛋白表达降低(均P<0.01),Ki67蛋白表达降低(P<0.01),且均呈浓度依赖性。结论秦皮甲素可能通过FBI-1调控细胞周期相关的p53-p21通路,影响DNA复制,从而抑制三阴性乳腺癌细胞的增殖。
Objective To investigate the effect of esculin on the proliferation of triple negative breast cancer cells and its molecular mechanism.Methods MDA-MB-231 cells were treated with 28,56,112,225,450 and 900μmol/L of esculin for 24,48 and 72 h,respectively,and the cell viability was detected by cell counting kit 8(CCK-8)assay.In addition,MDA-MB-231 cells were treated with 0,225,450 and 900μmol/L of esculin for 48 h.And then the changes in cell morphology were observed by inverted microscope.The clone-forming ability was detected by colony formation assay.The mRNA expression levels of FBI-1,p53 and p21 were detected using real-time fluorescence quantitative polymerase chain reaction.The protein expression levels of FBI-1,p53,p21 and Ki67 were detected by western blot.Results Compared with the blank control group,the cell viability of MDA-MB-231 cells that treated with esculin significantly decreased in a dose-dependent and time-dependent manners.After treatment with esculin,MDA-MB-231 cells shrunk,flattened,adhered poorly to the culture dish and the cell spacing became larger.Meanwhile,shedding and incomplete cells appeared,of which 900μmol/L of esculin treatment group showed the most dramatic changes.In addition,the colony formation ratios were decreased to(77.18±5.13)%,(65.94±4.98)%and(45.92±3.70)%in the 225,450 and 900μmol/L of esculin treatment groups compared with blank control,respectively(P<0.01).Furthermore,the mRNA and protein expressions of FBI-1 increased,while the levels of p53 and p21 mRNA and protein,as well as the protein expression of Ki67 decreased in a concentration-dependent manner(P<0.01).Conclusion Esculin may regulate cell cycle-related p53-p21 pathway via FBI-1 mediated DNA replication,thus inhibit the proliferation of triple negative breast cancer cells.
作者
莫淼
陈茂剑
黄怡
蒋玮
覃庆洪
梁至洁
杨伟萍
韦长元
Mo Miao;Chen Maojian;Huang Yi;Jiang Wei;Qin Qinghong;Liang Zhijie;Yang Weiping;Wei Changyuan(Department of Breast Surgery,Guangxi Medical University Cancer Hospital,Nanning 530021,China;Department of Experimental Research,Guangxi Medical University Cancer Hospital,Nanning 530021,China;Department of Respiratory Oncology,Guangxi Medical University Cancer Hospital,Nanning 530021,China;Breast Center,the Fifth Affiliated Hospital of Guangxi Medical University,Nanning 530021,China;Department of Ultrasound,Guangxi Medical University Cancer Hospital,Nanning 530021,China)
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2020年第8期629-634,共6页
Chinese Journal of Oncology
基金
广西自然科学基金(2015GXNSFAA139204)
广西研究生教育创新计划(YCSW2017108,YCBZ2018041)。
