四君子汤提取物对人乳腺癌MDA-MB-468细胞生长的抑制作用  

Inhibition of Sijunzi Decoction extract on human breast cancer cells MDA-MB-468

作　　者：李朵璐[1] 张待娣 杨盟 肖冉 段震峰[1,2] LI Duo-lu;ZHANG Dai-di;YANG Meng;XIAO Ran;DUAN Zhen-feng(Henan Key Laboratory for Precision Clinical Pharmacy,Department of Pharmacy,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Sarcoma Biology Laboratory,UCLA Orthopaedic Surgery,Los Angeles 90095,USA)

机构地区：[1]郑州大学第一附属医院药学部河南省精准临床药学重点实验室,河南郑州450052 [2]美国加州大学洛杉矶分校大卫格芬医学院骨科实验室,美国洛杉矶90095

出　　处：《中草药》2020年第4期1037-1043,共7页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金面上项目(31670895);河南省科技发展计划项目(182102410007)。

摘　　要：目的探究四君子汤提取物(Sijunzi Decoction extract,SDE)对人三阴性乳腺癌MDA-MB-468细胞生长的作用。方法采用不同质量浓度SDE作用于MDA-MB-468细胞,CCK-8实验和细胞划痕实验检测SDE对细胞增殖和迁移能力的影响;克隆形成实验检测SDE对细胞集落形成能力的影响;Hoechst33342染色技术和流式细胞术(FCM)检测SDE对细胞凋亡和周期的影响;Western blotting技术检测SDE对信号传导及转录激活蛋白3(STAT3)表达的影响。结果与对照组比较,SDE对MDA-MB-468细胞有一定的抑制作用(P<0.05),且呈质量浓度和时间依赖性。克隆形成实验结果表明SDE能够抑制细胞克隆形成。细胞迁移实验结果显示,SDE中、高质量浓度能够明显减弱细胞划痕愈合能力(P<0.001)。FCM检测凋亡结果显示,SDE各质量浓度组细胞早期凋亡率和总凋亡率逐渐上升,呈质量浓度依赖性,且中、高质量浓度的SDE能显著诱导细胞凋亡(P<0.01、0.001)。SDE能够影响细胞周期,使G2期细胞显著减少(P<0.01)。Westernblotting结果显示,SDE处理细胞后,STAT3蛋白表达水平显著降低。结论SDE能够抑制MDA-MB-468细胞的增殖和克隆形成,促进其凋亡,使G2期细胞减少。其机制可能与调控STAT3通路有关。Objective To investigate the effect of Sijunzi Decoction extract(SDE) on the growth of human triple negative breast cancer(TNBC) cell line MDA-MB-468. Methods MDA-MB-468 cells were treated with different concentrations of SDE. The effect of SDE on the proliferation and migration of the cells were detected by CCK-8 assay and the cell wound healing assay. The colony formation assay was performed to analyze the effect on the ability of colony formation of the cells with SDE. Hoechst 33342 staining technique and flow cytometry(FCM) were used to detect apoptosis and cell cycle of the cells. Western blotting was used to detect the expression levels of STAT3, which was related with the proliferation and apoptosis of cells. Results Compared with the control group, SDE had a certain inhibitory effect on MDA-MB-468 cells(P < 0.05), and it was dependent on the concentration and time. Cloning formation experiments showed that SDE inhibited the clonality of the cells. The cell migration experiment showed that the wound healing ability of the cells could be weakened by the extract with the medium and high dosage(P < 0.001). The results of FCM showed that the apoptosis rate of all SDE dosage increased gradually in a dose-dependent manner. And SDE with the medium and high dosage induced apoptosis of the cells significantly(P < 0.01 and 0.001). Cell cycle was affected by SDE with the obvious reduction of the cells in G2 phase(P < 0.01). The results of Western blotting showed that the expression level of STAT3 was decreased significantly. Conclusion SDE inhibited the proliferation and clonal formation of MDA-MB-468 cells, inhibited migration, promoted apoptosis and decreased the cells of G2 phase. which may be related to the regulation of STAT3 pathway.

关 键 词：四君子汤 三阴性乳腺癌 增殖 凋亡 迁移 细胞周期 信号传导及转录激活蛋白3 

分 类 号：R285.5[医药卫生—中药学]