Graduate Institute of Chinese Medicine, China Medical University, 40402 Taichung, Taiwan, R.O.C.
Int J Oncol. 2013 Jul;43(1):338-48. doi: 10.3892/ijo.2013.1942. Epub 2013 May 16.
Liver cancer is the fifth most common cause of cancer death worldwide. The study of more effective anti-hepatoma drugs is urgently required. Bufalin has been isolated from a traditional Chinese medicine and possesses less toxicity to normal cells. However, it has been found to inhibit growth of cancer cells. In this study, we aimed to investigate the efficacy and mechanism of bufalin in Huh7, Hep3B and HA22T human hepatoma cells. The three cell lines were treated with bufalin, the proliferation was detected by WST-1 assay and cell cycle was detected by flow cytometry analysis. The results showed that bufalin inhibited the proliferation of hepatoma cells and regulated the hepatoma cell death program in a dose- and time-dependent manner without typical features of apoptosis. RT-PCR arrays were used to investigate the autophagy transcriptional response triggered by bufalin and 13 genes were altered and further confirmed by real-time PCR. The translation levels of selected genes were examined by western blot analysis to reveal the bufalin-induced autophagy cascade. Bufalin synergized with the JNK pathway to induce autophagy of hepatoma cells and is closely associated with the upregulation of TNF, BECN-1, MAPK and ATG8, together with the downregulation of Bcl-2 and Bid. Our study provided a multi-angle evaluation system for anti-hepatoma pharmacology for pre-clinical drug investigation. In this case, bufalin was capable of inducing hepatoma cell autophagy, suggesting a potential regimen for single or combined chemotherapy to overcome hepatoma in clinical practice.
肝癌是全球第五大常见癌症死因。迫切需要研究更有效的抗肝癌药物。蟾毒灵已从一种中药中分离出来，对正常细胞的毒性较小。然而，它已被发现能抑制癌细胞的生长。在这项研究中，我们旨在研究蟾毒灵在 Huh7、Hep3B 和 HA22T 人肝癌细胞中的疗效和机制。用蟾毒灵处理这三种细胞系，通过 WST-1 检测法检测细胞增殖，通过流式细胞术分析检测细胞周期。结果表明，蟾毒灵抑制肝癌细胞的增殖，并以剂量和时间依赖的方式调节肝癌细胞的死亡程序，而没有典型的凋亡特征。RT-PCR 阵列用于研究蟾毒灵触发的自噬转录反应，有 13 个基因发生改变，并通过实时 PCR 进一步证实。通过 Western blot 分析检查选定基因的翻译水平，以揭示蟾毒灵诱导的自噬级联。蟾毒灵与 JNK 通路协同诱导肝癌细胞自噬，与 TNF、BECN-1、MAPK 和 ATG8 的上调以及 Bcl-2 和 Bid 的下调密切相关。我们的研究为临床前药物研究提供了一个抗肝癌药理学的多角度评价体系。在这种情况下，蟾毒灵能够诱导肝癌细胞自噬，这表明在临床实践中，单一或联合化疗可能有治疗肝癌的潜力。
