摘要
探讨半夏泻心汤(Banxia Xiexin Decoction,BXD)对结肠癌细胞增殖、凋亡、侵袭及迁移的影响及作用机制。首先基于UPLC-MS/MS对半夏泻心汤入血成分进行鉴定,并利用HPLC技术进行含量检测,然后分别用不同浓度的BXD干预NCM460正常肠上皮细胞及HT29和HCT116结肠癌细胞,通过细胞增殖与活性检测-8(cell counting kit-8,CCK-8)法检测BXD对细胞活力的影响;通过流式细胞术检测BXD对细胞凋亡的影响,并采用Western blot法检测BXD对B细胞淋巴瘤/白血病-2(B-cell lymphoma/leukemia-2,Bcl-2)、B细胞淋巴瘤/白血病-2相关X蛋白(Bcl-2 associated X,Bax)表达的影响;通过细胞划痕实验检测BXD对细胞迁移能力的影响,Transwell侵袭法检测BXD对细胞侵袭的影响,并通过Western blot法检测BXD对上皮间质转化(epithelial-mesenchymal transition,EMT)相关蛋白如上皮型钙黏蛋白(epithelial cadherin,E-cadherin)、神经型钙黏蛋白(neural cadherin,N-cadherin)、波形蛋白(vimentin)表达的影响;采用Western blot及RT-qPCR技术检测BXD对聚(ADP-核糖)糖水解酶[poly(ADP-ribose)glycohydrolase,PARG]/聚(ADP-核糖)聚合酶1[poly(ADP-ribose)polymerase 1,PARP1]/核因子-κB p65(nuclear factor kappa-B p65,NF-κB p65)信号通路蛋白及相关mRNA表达水平的影响。结果表明,BXD干预后,HT29和HCT116结肠癌细胞增殖能力减弱,BXD可促进结肠癌细胞凋亡,促进Bcl-2蛋白表达,抑制Bax蛋白表达;同时,BXD可以抑制细胞迁移和侵袭,使E-cadherin蛋白表达升高,N-cadherin和vimentin蛋白表达降低;此外,BXD能抑制PARG、PARP1、NF-κB p65蛋白及其mRNA表达。综上,BXD可能通过介导PARG/PARP1/NF-κB信号通路从而抑制结肠癌细胞恶性表型。
This study aims to delve into the influences and underlying mechanisms of Banxia Xiexin Decoction(BXD)on the proliferation,apoptosis,invasion,and migration of colon cancer cells.Firstly,the components of BXD in blood were identified by UPLC-MS/MS,and subsequently the content of these components were determined by HPLC.Then,different concentrations of BXD were used to treat both the normal intestinal epithelial cells(NCM460)and the colon cancer cells(HT29 and HCT116).The cell viability and apoptosis were examined by the cell counting kit-8(CCK-8)and flow cytometry,respectively.Western blot was employed to determine the expression of the apoptosis regulators B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X(Bax).The cell wound healing assay and Transwell assay were employed to measure the cell migration and invasion,respectively.Additionally,Western blot was employed to determine the expression levels of epithelial-mesenchymal transition(EMT)-associated proteins,including epithelial cadherin(E-cadherin),neural cadherin(N-cadherin),and vimentin.The protein and mRNA levels of the factors in the poly(ADPribose)glycohydrolase(PARG)/poly(ADP-ribose)polymerase 1(PARP1)/nuclear factor kappa-B p65(NF-κB p65)signaling pathway were determined by Western blot and RT-qPCR,respectively.The results demonstrated that following BXD intervention,the proliferation of HT29 and HCT116 cells was significantly reduced.Furthermore,BXD promoted the apoptosis,enhanced the expression of Bcl-2,and suppressed the expression of Bax in colon cancer cells.At the same time,BXD suppressed the cell migration and invasion and augmented the expression of E-cadherin while diminishing the expression of N-cadherin and vimentin.In addition,BXD down-regulated the protein and mRNA levels of PARG,PARP1,and NF-κB p65.In conclusion,BXD may inhibit the malignant phenotypes of colon cancer cells by mediating the PARG/PARP1/NF-κB signaling pathway.
作者
黄渝清
王佳梅
赖恒周
肖冲
由凤鸣
旷歧轩
蒋义芳
HUANG Yu-qing;WANG Jia-mei;LAI Heng-zhou;XIAO Chong;YOU Feng-ming;KUANG Qi-xuan;JIANG Yi-fang(TCM Regulating Metabolic Diseases Key Laboratory of Sichuan Province,Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China;Cancer Institute,Chengdu University of Traditional Chinese Medicine,Chengdu 610072,China)
出处
《中国中药杂志》
北大核心
2025年第2期496-506,共11页
China Journal of Chinese Materia Medica
基金
四川省自然科学基金项目(2024NSFSC1849)
国家资助博士后研究人员计划项目(GZC20230334)。
