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.
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.
Zhongguo Zhong Yao Za Zhi. 2025 Jan;50(2):496-506. doi: 10.19540/j.cnki.cjcmm.20240923.304.
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(ADP-ribose) 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.
本研究旨在探讨半夏泻心汤（BXD）对结肠癌细胞增殖、凋亡、侵袭和迁移的影响及其潜在机制。首先，采用超高效液相色谱-串联质谱（UPLC-MS/MS）鉴定BXD的血中成分，随后用高效液相色谱（HPLC）测定这些成分的含量。然后，用不同浓度的BXD处理正常肠上皮细胞（NCM460）和结肠癌细胞（HT29和HCT116）。分别采用细胞计数试剂盒-8（CCK-8）和流式细胞术检测细胞活力和凋亡情况。采用蛋白质免疫印迹法检测凋亡调节因子B细胞淋巴瘤-2（Bcl-2）和Bcl-2相关X蛋白（Bax）的表达。分别采用细胞划痕愈合试验和Transwell试验检测细胞迁移和侵袭能力。此外，采用蛋白质免疫印迹法检测上皮-间质转化（EMT）相关蛋白，包括上皮钙黏蛋白（E-cadherin）、神经钙黏蛋白（N-cadherin）和波形蛋白的表达水平。分别采用蛋白质免疫印迹法和逆转录-定量聚合酶链反应（RT-qPCR）检测聚（ADP-核糖）糖苷水解酶（PARG）/聚（ADP-核糖）聚合酶1（PARP1）/核因子κB p65（NF-κB p65）信号通路相关因子的蛋白和mRNA水平。结果表明，BXD干预后，HT29和HCT116细胞的增殖明显降低。此外，BXD促进结肠癌细胞凋亡，增强Bcl-2表达，抑制Bax表达。同时，BXD抑制细胞迁移和侵袭，增加E-cadherin表达，降低N-cadherin和波形蛋白表达。此外，BXD下调PARG、PARP1和NF-κB p65的蛋白和mRNA水平。综上所述，BXD可能通过介导PARG/PARP1/NF-κB信号通路抑制结肠癌细胞的恶性表型。
