First Department of Internal Medicine, Mie University School of Medicine, Tsu, Japan.
Int J Oncol. 2013 Jan;42(1):101-8. doi: 10.3892/ijo.2012.1676. Epub 2012 Oct 24.
A multi-kinase inhibitor, sorafenib, was recently approved and is currently recommended for the treatment of advanced hepatocellular carcinoma (HCC). However, HCC treatment outcomes are still poor and necessitate improvement. Therefore, we investigated the influence of sorafenib in combination with each of cytotoxic chemotherapy agents, hypoxia or tumor necrosis factor (TNF)-related apoptosis‑inducing ligand (TRAIL), on cytotoxicity to determine which is the better adjuvant. Additive cytotoxicity of sorafenib to chemotherapy agents, hypoxia and TRAIL, to HCC cells was assessed using cell viability assay. Intracellular levels of anti-apoptotic proteins were determined using western blot analysis. Activation of Wnt/β-catenin signaling was assessed using a luciferase reporter gene assay. Sorafenib significantly and synergistically enhanced the cytotoxicity of TRAIL to HCC cells and 4',6-diamidino-2-phenylindole (DAPI) staining showed increased apoptosis among cells treated with sorafenib and TRAIL. This augmentation in cytotoxicity was derived from sorafenib-mediated downregulation of anti-apoptotic proteins. However, sorafenib did not enhance the cytotoxicity of chemotherapy agents (cisplatin, 5-FU or doxorubicin) or hypoxic treatment to HCC. Moreover, hypoxic treatment induced Wnt/β-catenin signaling activation. Our data showed that in combination TRAIL and sorafenib had a synergistic cytokilling effect on HCC cells and that this effect derived from sorafenib-mediated downregulation of anti-apoptotic proteins.
一种多激酶抑制剂，索拉非尼，最近被批准，目前被推荐用于治疗晚期肝细胞癌（HCC）。然而，HCC 的治疗效果仍然很差，需要改进。因此，我们研究了索拉非尼与细胞毒性化疗药物、缺氧或肿瘤坏死因子（TNF）相关凋亡诱导配体（TRAIL）联合使用对细胞毒性的影响，以确定哪种辅助治疗更好。使用细胞活力测定法评估索拉非尼与化疗药物、缺氧和 TRAIL 联合对 HCC 细胞的细胞毒性的增效作用。使用 Western blot 分析测定细胞内抗凋亡蛋白的水平。使用荧光素酶报告基因测定法评估 Wnt/β-catenin 信号转导的激活。索拉非尼显著协同增强 TRAIL 对 HCC 细胞的细胞毒性，并且 DAPI 染色显示在用索拉非尼和 TRAIL 处理的细胞中凋亡增加。这种细胞毒性的增强来自于索拉非尼介导的抗凋亡蛋白下调。然而，索拉非尼并没有增强化疗药物（顺铂、5-FU 或阿霉素）或缺氧处理对 HCC 的细胞毒性。此外，缺氧处理诱导了 Wnt/β-catenin 信号转导的激活。我们的数据表明，TRAIL 和索拉非尼联合使用对 HCC 细胞具有协同的细胞杀伤作用，这种作用源于索拉非尼介导的抗凋亡蛋白下调。
