Guangxi Key Laboratory for Bioactive Molecules Research and Evaluation & College of Pharmacy, Guangxi Medical University, 22 Shuangyong Road, Nanning, 530021, China; Key Laboratory of Longevity and Aging-related Diseases of Chinese Ministry of Education & Center for Translational Medicine, Guangxi Medical University, 22 Shuangyong Road, Nanning, 530021, China.
Guangxi Key Laboratory for Bioactive Molecules Research and Evaluation & College of Pharmacy, Guangxi Medical University, 22 Shuangyong Road, Nanning, 530021, China.
J Ethnopharmacol. 2022 Sep 15;295:115381. doi: 10.1016/j.jep.2022.115381. Epub 2022 May 17.
Marsdenia tenacissima injection (MTE), a traditional Chinese medical injection extracted from the rattan of Marsdenia tenacissima (Roxb.) Moon, has been approved for clinical use in China as an adjuvant therapeutic agent in multiple cancers, including esophageal cancer, gastric cancer, lung cancer, and liver cancer. However, the activity and mechanism of MTE on prostate cancer (PCa) remain to be defined.
To investigate the activity and the underlying mechanism of MTE in the treatment of PCa.
The component characterization of MTE was analyzed by HPLC-CAD-QTOF-MS/MS technology. Cell Counting Kit-8 (CCK-8) assay was used to assess PCa cell proliferation. Colony formation assay was applied to detect the clonogenic ability of the cells. MetaboAnalyst5.0 database was employed to analyze the altered metabolites of PC3 cells treated with MTE obtained by UPLC-QTOF-MS/MS. Combined with metabolomics analysis and network pharmacology, we predicted the potential targets, which further were verified by Western Blot, RT-qPCR, and Immunohistochemistry assays. Finally, SeeSAR software was applied to predict the potential active components of MTE against PCa.
A total of 21 components in MTE were confirmed by HPLC-CAD-QTOF-MS/MS analysis. MTE inhibited the proliferation and colony formation of PCa cells. A total of 20 metabolites closely related to glycerophospholipid metabolism, glycolysis/gluconeogenesis, and tricarboxylic acid (TCA) cycle were significantly changed in PC3 cells treated with MTE. The network pharmacology analysis revealed that MTE suppressed the growth of PC3 cells might by regulating the ErbB2-GSK3β-HIF1α signaling axis. Furthermore, we also confirmed that stimulation of MTE significantly inhibited the phosphorylation of ErbB2 at Tyr877 and the activities of its downstream signal transducers (GSK3β and HIF1α) in PCa, as well as the mRNA levels of critical factors (IDH2, LDHA, and HIF1A) in the tricarboxylic acid (TCA) cycle. Molecular docking further suggested that Tenacissimoside E, cryptochlorogenic acid, and scopoletin might be the active ingredients of MTE for PCa treatment.
This study proposed that MTE exerts a potential anti-tumor effect in PCa through inhibiting ErbB2-GSK3β-HIF1α signaling axis, which may be related to the TCA cycle.
从天仙藤（Marsdenia tenacissima （Roxb.） Moon）藤茎中提取的中药注射液马兜铃酸 tenacissima 注射液（MTE）已在中国被批准用于临床，作为多种癌症（包括食管癌、胃癌、肺癌和肝癌）的辅助治疗药物。然而，MTE 对前列腺癌（PCa）的活性和机制仍有待确定。
研究 MTE 治疗 PCa 的活性及其作用机制。
采用高效液相色谱-串联飞行时间质谱联用技术（HPLC-CAD-QTOF-MS/MS）分析 MTE 的成分特征。使用细胞计数试剂盒-8（CCK-8）测定法评估 PCa 细胞增殖。应用集落形成实验检测细胞的克隆形成能力。采用代谢组学分析和网络药理学，我们预测了用 UPLC-QTOF-MS/MS 处理后的 PC3 细胞中受 MTE 影响的代谢物，并进一步通过 Western Blot、RT-qPCR 和免疫组织化学检测进行了验证。最后，使用 SeeSAR 软件预测 MTE 治疗 PCa 的潜在活性成分。
通过 HPLC-CAD-QTOF-MS/MS 分析，共确定了 MTE 中的 21 种成分。MTE 抑制了 PCa 细胞的增殖和集落形成。MTE 处理后的 PC3 细胞中，与甘油磷脂代谢、糖酵解/糖异生和三羧酸（TCA）循环密切相关的 20 种代谢物发生了显著变化。网络药理学分析表明，MTE 可能通过调节 ErbB2-GSK3β-HIF1α 信号轴来抑制 PC3 细胞的生长。此外，我们还证实，MTE 的刺激显著抑制了 PCa 中 ErbB2 在 Tyr877 处的磷酸化以及其下游信号转导物（GSK3β 和 HIF1α）的活性，以及三羧酸（TCA）循环中的关键因子（IDH2、LDHA 和 HIF1A）的 mRNA 水平。分子对接进一步表明，Tenacissimoside E、隐绿原酸和东莨菪内酯可能是 MTE 治疗 PCa 的活性成分。
本研究提出 MTE 通过抑制 ErbB2-GSK3β-HIF1α 信号轴在 PCa 中发挥潜在的抗肿瘤作用，这可能与 TCA 循环有关。
