摘要
目的:观察芪蓝方对人前列腺癌DU145细胞增殖和凋亡的影响,并探讨其作用机制。方法:以DU145细胞为研究对象,通过观察不同浓度芪蓝方组(400、200、100、50、25、12.5、6.25、3.125、1.56、0μg/ml)细胞形态变化,筛选出高、中、低浓度应用于后续实验,并采用CCK-8法检测DU145细胞增殖,流式细胞术检测DU145细胞周期、凋亡情况,Western印迹检测DU145细胞中细胞周期、凋亡相关蛋白Cyclin D1、Bax、Bcl-2、Cleaved-Caspase 3的表达。结果:筛选结果显示100、200、400μg/ml浓度的芪蓝方均能显著抑制DU145细胞的生长、降低轮廓清晰度和贴壁能力,且200、400μg/ml浓度的芪蓝方可显著降低DU145细胞活力,故确定高中低浓度为400、200、100μg/ml,并用于后续实验研究。与空白组比较,G2期各浓度组DU145细胞数显著增加(P<0.01),而S期高、中浓度组细胞数显著下降(P<0.01、P<0.05);与空白对照组相比,芪蓝方各组DU145细胞总凋亡数均显著增高(P<0.01),高浓度组Cyclin D1表达显著降低,高、中浓度组Bcl-2表达明显降低,Bax和Cleaved-Caspase 3表达水平明显上升(P均<0.01)。结论:芪蓝方能够抑制前列腺癌DU145细胞的细胞增殖,促进其细胞凋亡,其机制可能与下调细胞周期相关蛋白Cyclin D1表达,破坏Bax/Bcl-2平衡,上调Cleaved-Caspase 3表达相关。
Objective:To investigate the effects of different concentrations of Qilan Prescription(QLP)on the proliferation and apoptosis of human PCa DU145 cells and its underlying mechanism.Methods:We treated human PCa DU145 cells with QLP at 400,200,100,50,25,12.5,6.25,3.125 or 1.56μg/ml for 24,48 and 72 hours respectively.Then we observed the morphological changes of the cells,examined their viability by CCK-8 assay,detected their cell cycle and apoptosis by flow cytometry,and determined the protein expressions of cyclin D1,Bax,Bcl-2 and cleaved-caspase 3 in the DU145 cells by Western blot,followed by comparison of the parameters with those obtained from the blank control group.Results:QLP significantly inhibited the growth,reduced the contour clarity and adhesion ability of the DU145 cells at the concentrations of 100,200 and 400μg/ml,and markedly decreased the activity of the cells at 200 and 400μg/ml,most significantly at 400μg/ml.The number of the G2-phase DU145 cells was dramatically increased in all the concentration groups(P<0.01),so was the total number of apoptotic DU145 cells(P<0.01),while that of the S-phase cells remarkably decreased in the 400μg/ml QLP(P<0.01)and 200μg/ml QLP(P<0.05)groups.The expression of the cyclin D1 protein was significantly down-regulated in the 400μg/ml QLP group(P<0.01).That of Bcl-2 was also down-regulated(P<0.01)while those of Bax and cleaved-caspase 3 up-regulated in the 400 and 200μg/ml QLP groups(P<0.01).Conclusion:QLP can inhibit the proliferation and promote the apoptosis of human PCa DU145 cells,which may be associated with its effects of down-regulating the expression of the cell cycle-related protein cyclin D1,disrupting the Bax-Bcl-2 balance,and up-regulating the expression of cleaved-caspase 3.
作者
周静
原凡
骆春梅
朱坤
刘柏言
昌玥
尤耀东
ZHOU Jing;YUAN Fan;LUO Chun-mei;ZHU Kun;LIU Bo-yan;CHANG Yue;YOU Yao-dong(School of Clinical Medicine,Chengdu University of Chinese Medicine,Chengdu,Sichuan 610075,China;Department of Urology and Andrology,The Affiliated Hospital of Chengdu University of Chinese Medicine,Chengdu,Sichuan 610072,China;Key Laboratory of Traditional Chinese Medicine Regulation of Metabolic Diseases,Chengdu,Sichuan 610072,China)
出处
《中华男科学杂志》
CAS
CSCD
北大核心
2023年第3期255-263,共9页
National Journal of Andrology
基金
国家自然科学基金面上项目(81973866)
四川省科技厅自然科学基金项目(2022NSFSC0684)
四川省中医药管理局科学技术研究专项(2021ZD016)
成都中医药大学“杏林学者”学科人才科研提升计划研究专项(QNXZ2019020)。
