摘要
目的:探究薯蓣丸联合紫杉醇抑制人三阴性乳腺癌MDA-MB-231细胞上皮间质转化(EMT)的作用和机制。方法:运用血清药理学方法制备SD大鼠空白血清和薯蓣丸含药血清,体外培养MDA-MB-231细胞。实验将其分组为空白组、紫杉醇组、薯蓣丸联合紫杉醇组三组。运用CCK8实验检测MDA-MB-231细胞的增殖情况;Transwell小室实验法检测细胞的侵袭、迁移能力变化;Western blot检测上皮间质转化相关分子标志物E-cadherin、Vimentin、N-cadherin、MMP2和PI3K/Akt/mTOR信号通路相关蛋白的表达;RT-qPCR检测E-cadherin、Vimentin、N-cadherin、MMP2 mRNA的表达。结果:与空白组相比,紫杉醇组和薯蓣丸联合紫杉醇组可显著抑制细胞的增殖、侵袭和迁移(P<0.01);与紫杉醇组相比,薯蓣丸联合紫杉醇组对细胞增殖、侵袭和迁移的抑制能力增强(P<0.01)。与空白组相比,紫杉醇组和薯蓣丸联合紫杉醇组细胞中E-cadherin蛋白表达上调,PI3K、Akt、mTOR、N-cadherin、Vimentin蛋白表达下调(P<0.05,P<0.01);与紫杉醇组相比,薯蓣丸联合紫杉醇组细胞中E-cadherin蛋白表达上调,Vimentin、MMP2、PI3K、Akt、p-Akt、mTOR、p-mTOR蛋白表达下调(P<0.05,P<0.01)。与空白组比较,紫杉醇组和薯蓣丸联合紫杉醇组均能上调E-cadherin,下调Vimentin、MMP2 mRNA的表达(P<0.01);与紫杉醇组比较,薯蓣丸联合紫杉醇组上调E-cadherin,下调N-cadherin、MMP2 mRNA表达能力增强(P<0.05,P<0.01)。结论:薯蓣丸联合紫杉醇可有效抑制三阴性乳腺癌细胞的侵袭、迁移和EMT进程,且效果优于紫杉醇单用,其机制可能与PI3K/Akt/mTOR信号通路受抑制相关。
Objective:To investigate the effect and mechanism of Dioscorea pill combined with paclitaxel on Epithelial-mesenchymal transition(EMT)in human triple-negative breast cancer MDA-MB-231 cells.Methods:SD rat serum and dioscorea zingiberensis pill containing serum were prepared by serum pharmacology method,and MDA-MB-231 cells were cultured in vitro.The experimental groups were blank group,paclitaxel group and Dioscorea pill combined with paclitaxel group.CCK8 assay was used to detect cell proliferation.The invasion and migration of cells were detected by Transwell assay.Western blot was used to detect the expression of epithelial-mesenchymal transition related molecular markers E-cadherin,Vimentin,N-cadherin,MMP2 and PI3K/Akt/mTOR signaling pathway related proteins.RT-qPCR was used to detect the mRNA expression of E-cadherin,Vimentin,N-cadherin and matrix metalloproteinase 2(MMP2).Results:Compared with the control group,the proliferation,invasion and migration of MDA-MB-231 cells were significantly inhibited in the paclitaxel group and the dioscin pill combined with paclitaxel group(P<0.01).Compared with the paclitaxel group,the inhibition effect of dioscorea pill combined with paclitaxel on cell proliferation,invasion and migration was significantly enhanced(P<0.01).Compared with the blank group,the expression of E-cadherin protein was up-regulated in paclitaxel group and dioscorea pill combined with paclitaxel group,while the expression of N-cadherin,Vimentin,PI3K,Akt and mTOR protein was down-regulated(P<0.05,P<0.01).Compared with paclitaxel group,the expression of E-cadherin protein in dioscorea pill combined with paclitaxel group was up-regulated,and the expression of Vimentin,MMP2,PI3K,Akt,p-Akt,mTOR and p-mTOR protein was down-regulated(P<0.05,P<0.01).Compared with the control group,the expression of E-cadherin in paclitaxel group and diosgenin pill combined with paclitaxel group was up-regulated,and the mRNA expressions of Vimentin and MMP2 were down-regulated(P<0.01).Compared with the paclitaxel group,the Dioscorea pill combined with paclitaxel group had a significant increase in the mRNA expression of E-cadherin and a significant decrease in the mrna expression of N-cadherin and MMP2(P<0.05,P<0.01).Conclusion:Dioscorea pill combined with paclitaxel can inhibit the migration,invasion and EMT process of triple-negative breast cancer MDA-MB-231 cells,and the effect is better than that of paclitaxel alone.The mechanism may be related to the down-regulation of PI3K/Akt/mTOR signaling pathway.
作者
骆小珊
谢甦
朱曼荆
黄雅珍
朱久龙
冯豆豆
谢青
蒙雁云
凌湘力
LUO Xiaoshan;XIE Su;ZHU Manjing;HUANG Yazhen;ZHU Jiulong;FENG Doudou;XIE Qing;MENG Yanyun;LING Xiangli(Guizhou Medical University,Guiyang 550004,China)
出处
《陕西中医》
CAS
2023年第9期1163-1168,共6页
Shaanxi Journal of Traditional Chinese Medicine
基金
国家自然科学基金资助项目(81960834)
贵州省科技计划项目[黔科合基础-ZK(2022)一般442]
贵州省中医药管理局中医药、民族医药科学技术研究课题(QZYY-2023-001)
国家中医药管理局第二届全国名中医凌湘力传承工作室项目[国中医药人教函(2021)202号]。
