芒柄花苷通过下调Cyclin D表达抑制乳腺癌细胞生长的机制研究  

Study on the mechanism of Ononin in inhibiting breast cancer growth by down-regulating Cyclin D

作　　者：黄秋艳[1] 林庆中[1] 吴秀凤[1] 黄丽英[1] 黄志坚[1] 叶文飞[1] HUANG Qiu-yan;LIN Qing-zhong;WU Xiu-feng;HUANG Li-ying;HUANG Zhi-jian;YE Wen-fei(Department of Breast Surgical Oncology,Fujian Medical University Cancer Hospital,Fujian Cancer Hospital,Fuzhou 350014,Fuyjian Province,China)

机构地区：[1]福建医科大学附属肿瘤医院/福建省肿瘤医院乳腺肿瘤外科,福建福州350014

出　　处：《中国临床药理学杂志》2022年第17期1999-2003,共5页The Chinese Journal of Clinical Pharmacology

基　　金：福建省科技厅卫生联合资金资助项目(2018J01264)。

摘　　要：目的 探究黄酮化合物芒柄花苷对乳腺癌生长的影响及其确切机制。方法 取对数生长期人乳腺癌MCF-7、MDA-MB-231、MDA-MB-468细胞,3种细胞均随机分为对照组、低剂量实验组、中剂量实验组、高剂量实验组、Cyclin D沉默组和高剂量实验组+Cyclin D沉默组。3种细胞的低、中、高剂量实验组均分别以浓度为20、40、80μmol·L^(-1)的芒柄花苷处理24 h,对照组使用等量二甲基亚砜处理。Cyclin D沉默组将3种细胞分别转染Cyclin D siRNA 24 h;高剂量实验组+Cyclin D沉默组在实验开始前24 h,3种细胞分别转染Cyclin D siRNA。实验0 h时用80μmol·L^(-1)的芒柄花苷处理24 h。用细胞计数试剂盒-8(CCK-8)实验检测细胞增殖能力;用碘化丙啶单染细胞周期实验检测细胞周期进程;用BrdU掺入实验来检测细胞DNA复制活性;用蛋白质印迹法检测细胞中Cyclin D的表达。结果 在MCF-7细胞中,对照组和低、中、高剂量实验组细胞的增殖抑制率分别为(0.00±8.24)%,(7.20±16.81)%,(38.87±10.19)%和(65.60±5.39)%;这4组的G0/G1期比例分别为(33.87±0.68)%,(38.90±2.84)%,(44.63±3.45)%和(58.53±3.10)%;这4组的DNA复制比例分别为(100.00±6.23)%,(85.29±13.87)%,(56.67±10.26)%和(39.93±4.36)%;这4组的Cyclin D蛋白表达水平分别为1.00±0.14,0.84±0.19,0.66±0.13和0.43±0.05。在MCF-7细胞中,高剂量实验组、Cyclin D沉默组和高剂量实验组+Cyclin D沉默组细胞的增殖抑制率分别为(55.75±4.77)%,(22.65±10.31)%和(29.05±12.97)%。MDA-MB-231和MDA-MB-468细胞上述指标的变化趋势与MCF-7细胞相同。3种细胞的上述指标,中、高剂量实验组分别与对照组比较,差异均有统计学意义(均P<0.05)。结论 芒柄花苷可显著抑制人乳腺癌细胞增殖,使细胞周期阻滞于G0/G1期,抑制DNA复制活性,其机制可能与下调Cyclin D蛋白表达相关。Objective To investigate the effect of Ononin on breast cancer growth and its exact mechanism.Methods Human breast cancer MCF-7,MDA-MB-231 and MDA-MB-468 cells at logarithmic growth stage were randomly divided into control group,Ononin-L group,Ononin-M group,Ononin-H group,Cyclin D knockdown group and Ononin-H+Cyclin D knockdown group.The Ononin-L,-M,-H groups were treated with Ononin at the concentration of 20,40 and 80 μmol·L^(-1) for 24 h,respectively,while the control group was treated with the same concentration of dimethyl sulfoxide.Cyclin D knockdown group was transfected with Cyclin D si RNA for 24 h,and Ononin-H + Cyclin D knockdown group was transfected with Cyclin D si RNA for 24 h before the experiment.At 0 h of experiment,Ononin was treated with 80 μmol·L-1for 24 h.Cell proliferation was detected by cell counting kit 8( CCK-8).Cell cycle progression was detected by propidium iodide( PI) staining assay,and DNA replication activity was detected by Brd U incorporation assay.Cyclin D expression was detected by Western blot assay.Results In MCF-7 cells,the proliferation inhibition rates of control and Ononin-L,-M,-H groups were( 0.00 ± 8.24) %,( 7.20 ± 16.81) %,( 38.87 ± 10.19) % and( 65.60 ± 5.39) %,respectively;the proportion of G0/G1 phase in four groups were( 33.87 ± 0.68) %,( 38.90 ± 2.84) %,( 44.63 ± 3.45) % and( 58.53 ± 3.10) %;the proportion of DNA replication in four groups were( 100.00 ± 6.23) %,( 85.29 ± 13.87) %,( 56.67 ± 10.26) % and( 39.93 ± 4.36) %,respectively;the relative expression levels of Cyclin D protein in four groups were 1.00 ± 0.14,0.84 ± 0.19,0.66 ± 0.13 and 0.43 ± 0.05,respectively.In MCF-7 cells,the proliferation inhibition rates of Ononin-H group,Cyclin D knockdown group and Ononin-H + Cyclin D knockdown group were( 55.75 ± 4.77) %,( 22.65 ± 10.31) % and( 29.05 ± 12.97) %,respectively.MDA-MB-231 and MDA-MB-468 cells showed the same change trend as MCF-7 cells.The above indexes of 3 kinds of cells,the differences between control group and the Ononin-L,-H

关 键 词：芒柄花苷 乳腺癌 细胞增殖 Cyclin D蛋白 

分 类 号：R979.1[医药卫生—药品]