阳和汤对三阴性乳腺癌细胞MDA-MB-231凋亡Egr1/p21信号通路的影响  

Effect of Yanghetang on Apoptosis of Triple Negative Breast Cancer Cell Line MDA-MB-231 Based on Egr1/p21 Signaling Pathway

作　　者：李康乐 杨小蒨 张愉 杨硕 朱中博 窦建卫[1] LI Kang-le;YANG Xiao-qian;ZHANG Yu;YANG Shuo;ZHU Zhong-bo;DOU Jian-wei(College of Pharmacy,Xian Jiaotong University,Xian 710061,China;The Second Affiliated Hospital of XiSi Jiaotong University,Xidn 710004,China;College of Basic Medical Sciences,Gansu University of Chinese Medicine,Lanzhou 730000,China)

机构地区：[1]西安交通大学药学院,西安710061 [2]西安交通大学第二附属医院,西安710004 [3]甘肃中医药大学基础医学院,兰州730000

出　　处：《中国实验方剂学杂志》2020年第8期62-67,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81573983,81873311);陕西省中医药管理局科研课题项目(2019-ZZ-JC029);西安市中医药科研项目(SZJ201709);西安市中医院课题项目(YJ201915)。

摘　　要：目的:以早期生长反应基因1(Egr1)/细胞周期蛋白依赖性抑制剂(p21)信号通路为基础,探讨古方阳和汤含药血清对人三阴性乳腺癌细胞MDA-MB-231凋亡的影响。方法:制备阳和汤大鼠含药血清,体外培养三阴性乳腺癌MDA-MB-231细胞。设空白组,阳和汤高、中、低剂量组(23.4,11.7,5.85 g·kg^-1),分别干预MDA-MB-231细胞24,48,72 h,细胞增殖检测(CCK-8)法检测各组细胞增殖情况。设空白组,阳和汤高、中、低剂量组(23.4,11.7,5.85 g·kg^-1),干预MDA-MB-231细胞48 h,采用流式细胞仪检测各组细胞凋亡情况以及细胞周期分布情况;实时荧光定量聚合酶链式反应(Real-time PCR)检测各组细胞Egr1,p21 mRNA的表达情况;蛋白免疫印迹法(Western blot)检测各组细胞Egr1,p21蛋白的表达情况。结果:阳和汤干预MDA-MB-231细胞24,48,72 h,与空白组相比,阳和汤高、中剂量组能显著抑制MDA-MB-231细胞增殖(P<0.01)。阳和汤干预MDA-MB-231细胞48 h,与空白组相比,阳和汤高、中剂量组细胞的凋亡率明显增加(P<0.05,P<0.01);阳和汤高、中剂量组细胞周期G0/G1期所占比例降低,G2/M期所占比例升高(P<0.05,P<0.01);阳和汤高、中剂量组细胞Egr1,p21mRNA和蛋白表达均增加(P<0.05,P<0.01)。结论:阳和汤可激活MDA-MB-231细胞中Egr1/p21信号通路,增加Egr1,p21的表达,引起G2/M细胞周期阻滞,从而诱导MDA-MB-231细胞凋亡。Objective:To investigate the effect of ancient recipe Yanghetang and its drug-contained serum on apoptosis of triple negative breast cancer cell line MDA-MB-231 based on the signal pathway of early growth response gene 1(Egr1)/cyclin dependent inhibitor(p21).Method:The drug-containing serum of Yanghetang was prepared from rats,and MDA-MB-231 cells were cultured in vitro.The blank control group was set,and MDAMB-231 cells in Yanghetang high,middle,and low dose groups(23.4,11.7,5.85 g·kg^-1)were intervened for 24,48,72 h,respectively.After that,the cell counting kit-8(CCK-8)method was used to detect the cell proliferation of each group.The blank control group was set,while MDA-MB-231 cells in Yanghetang high,middle,and low dose groups(23.4,11.7,5.85 g·kg^-1)were treated for 48 h,and then flow cytometry was used to detect the apoptosis of each group and the distribution of cell cycle.The expression of Egr1 and p21 m RNA in each group was detected by quantificational Real-time polymerase chain reaction(Real-time PCR),while the expression of Egr1 and p21 protein in each group was detected by Western blot.Result:After MDA-MB-231 cells were intervened by Yanghetang for 24,48,72 h,MDA-MB-231 cell proliferation was significantly inhibited in Yanghetang high and middle dose groups as compared with the blank control group(P<0.01).After MDA-MB-231 cells were intervened by Yanghetang for 48 h,the apoptosis was significantly increased in Yanghetang high and middle dose groups as compared with the blank control group(P<0.05,P<0.01).In the Yanghetang high,middle dose groups,the proportion of cell cycle G0/G1 phase decreased,and the proportion of G2/M phase increased(P<0.05,P<0.01).The m RNA and protein expressions of Egr1,p21 were increased in Yanghetang high and middle groups(P<0.05,P<0.01).Conclusion:Yanghetang can activate Egr1/p21 signaling pathway in MDA-MB-231 cells,increase the expression of Egr1 and p21,and cause G2/M cell cycle arrest,thereby inducing apoptosis of MDA-MB-231 cells.

关 键 词：三阴性乳腺癌 MDA-MB-231细胞 阳和汤 早期生长反应基因1(Egr1) 细胞周期蛋白依赖性抑制剂(p21) 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]