State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Carcinogenesis and Intervention, China Pharmaceutical University, Nanjing, China.
Mol Pharmacol. 2012 Nov;82(5):958-71. doi: 10.1124/mol.112.079848. Epub 2012 Aug 21.
Tumor necrosis factor-α (TNF-α) is being used as an antineoplastic agent in treatment regimens of patients with locally advanced solid tumors, but TNF-α alone is only marginally active. In clinical use, it is usually combined with other chemical agents to increase its tumor response rate. Our previous studies reported that LYG-202 (5-hydroxy-8-methoxy-7-(4-(4-methylpiperazin-1-yl)butoxy)-2-phenyl-4H-chromen-4-one), a synthesized flavonoid with a piperazine substitution, has antiproliferative, antiangiogenic, and proapoptotic activities in multiple cancer cell lines. Here we evaluated the antineoplastic effect of TNF-α and analyzed the mechanism underlying its combination with LYG-202. Our results indicated that LYG-202 significantly increased the cytostatic and proapoptotic activity of TNF-α in HepG2 cells and heightened the protein level of apoptosis-related genes including caspase-3, caspase-8/9, cleaved poly(ADP-ribose) polymerase, and Bid. The fact that LYG-202 had a fitness score similar to that of the casein kinase 2 (CK2) inhibitor naphthyridine-8-carboxylate (CX-4945) implied to us that it may serve as a potential candidate for CK2 inhibitor, and the kinase activity assay suggested that LYG-202 significantly inhibited CK2 activity. Moreover, the electrophoretic mobility shift assay and luciferase assay showed that LYG-202 blocked the TNF-α-induced nuclear factor-κB (NF-κB) survival signaling pathway primarily by inactivating protein kinase CK2. In murine xenograft models, we also found that LYG-202 enhanced TNF-α antineoplastic activity and inhibited CK2 activity and NF-κB-regulated antiapoptotic gene expression. All these results showed that LYG-202 enhanced TNF-α-induced apoptosis by attenuating the CK2-dependent NF-κB pathway and probably is a promising agent in combination with TNF-α.
肿瘤坏死因子-α（TNF-α）被用作局部晚期实体瘤患者治疗方案中的抗肿瘤药物，但 TNF-α 单独使用时活性较低。在临床应用中，它通常与其他化学药物联合使用以提高其肿瘤反应率。我们之前的研究报告称，LYG-202（5-羟基-8-甲氧基-7-（4-（4-甲基哌嗪-1-基）丁氧基）-2-苯基-4H-色烯-4-酮），一种具有哌嗪取代基的合成黄酮类化合物，在多种癌细胞系中具有抗增殖、抗血管生成和促凋亡活性。在这里，我们评估了 TNF-α 的抗肿瘤作用，并分析了其与 LYG-202 联合的作用机制。我们的结果表明，LYG-202 显著增强了 TNF-α 在 HepG2 细胞中的细胞生长抑制和促凋亡活性，并提高了包括 caspase-3、caspase-8/9、cleaved poly（ADP-ribose）polymerase 和 Bid 在内的凋亡相关基因的蛋白水平。LYG-202 的拟合评分与酪蛋白激酶 2（CK2）抑制剂萘啶-8-羧酸（CX-4945）相似的事实表明，它可能是 CK2 抑制剂的潜在候选药物，激酶活性测定表明 LYG-202 可显著抑制 CK2 活性。此外，电泳迁移率变动分析和荧光素酶分析表明，LYG-202 通过使蛋白激酶 CK2 失活来阻断 TNF-α 诱导的核因子-κB（NF-κB）生存信号通路。在小鼠异种移植模型中，我们还发现 LYG-202 增强了 TNF-α 的抗肿瘤活性，抑制了 CK2 活性和 NF-κB 调节的抗凋亡基因表达。所有这些结果表明，LYG-202 通过减弱 CK2 依赖性 NF-κB 通路增强了 TNF-α 诱导的细胞凋亡，可能是与 TNF-α 联合应用的有前途的药物。
