Shanghai Key Laboratory of Compound Chinese Medicines, The Ministry of Education Key Laboratory for Standardization of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, P.R. China.
Institute of Interdisciplinary Integrative Medicine Sciences, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, P.R. China.
Int J Mol Med. 2021 Apr;47(4). doi: 10.3892/ijmm.2021.4882. Epub 2021 Feb 12.
Tumor necrosis factor‑α (TNF‑α) has different effects on apoptosis depending on activation or inactivation of the nuclear factor‑κB (NF‑κB) and epidermal growth factor receptor (EGFR) signaling pathways. Helichrysetin, a natural chalcone, inhibits NF‑κB nuclear translocation in mouse pancreatic β cells. The present study aimed to identify the effect of helichrysetin on activation of the NF‑κB and EGFR signaling pathways induced by TNF‑α, and the synergistic effect of helichrysetin and TNF‑α on apoptosis of HeLa and T98G cells. Cell proliferation was measured by Cell Counting Kit‑8 assay, while apoptosis was measured by Hoechst 33258 and Annexin V/PI staining. NF‑κB activity was detected by luciferase assay, protein expression was measured by western blotting and mRNA expression was detected by quantitative PCR assay. The results revealed that in HeLa and T98G cells helichrysetin blocked the increased phosphorylation of NF‑κB p65 induced by TNF‑α. Although helichrysetin alone decreased cell viability, helichrysetin and TNF‑α synergistically decreased cell viability. Helichrysetin, not TNF‑α, promoted apoptosis, while the combination of helichrysetin and TNF‑α synergistically increased apoptosis. In addition, helichrysetin and TNF‑α synergistically enhanced the activation of caspase‑3 and poly‑(ADP‑ribose)‑polymerase compared with helichrysetin alone. Helichrysetin inhibited the phosphorylation of transforming growth factor‑β activated kinase (TAK1), IκB kinase‑α/β (IKK‑α/β), NF‑κB p65 and EGFR induced by TNF‑α. Consistent with the inhibition of NF‑κB activation, the increased TNF‑α‑induced mRNA expression levels of TNF‑α, IL‑1β, CCL2, CCL5 and CXCL10 were significantly downregulated by helichrysetin. Therefore, helichrysetin and TNF‑α synergistically promoted apoptosis by inhibiting TAK1/IKK/NF‑κB and TAK1/EGFR signaling pathways in HeLa and T98G cells, indicating a potential therapeutic strategy for cancer.
肿瘤坏死因子-α（TNF-α）对细胞凋亡的影响取决于核因子-κB（NF-κB）和表皮生长因子受体（EGFR）信号通路的激活或失活。贺芹素是一种天然查尔酮，可抑制小鼠胰岛β细胞中 NF-κB 的核转位。本研究旨在确定贺芹素对 TNF-α诱导的 NF-κB 和 EGFR 信号通路激活的影响，以及贺芹素与 TNF-α协同作用对 HeLa 和 T98G 细胞凋亡的影响。通过 Cell Counting Kit-8 检测细胞增殖，通过 Hoechst 33258 和 Annexin V/PI 染色检测细胞凋亡。通过荧光素酶检测 NF-κB 活性，通过 Western blot 检测蛋白表达，通过 qPCR 检测 mRNA 表达。结果显示，在 HeLa 和 T98G 细胞中，贺芹素阻断了 TNF-α诱导的 NF-κB p65 磷酸化的增加。虽然贺芹素单独降低细胞活力，但贺芹素与 TNF-α协同降低细胞活力。贺芹素促进凋亡，而贺芹素与 TNF-α的组合协同增加凋亡。此外，与贺芹素单独作用相比，贺芹素与 TNF-α协同作用增强了 caspase-3 和多聚（ADP-核糖）聚合酶的激活。贺芹素抑制了 TNF-α诱导的转化生长因子-β激活激酶（TAK1）、IκB 激酶-α/β（IKK-α/β）、NF-κB p65 和 EGFR 的磷酸化。与 NF-κB 激活的抑制一致，贺芹素显著下调了 TNF-α诱导的 TNF-α、IL-1β、CCL2、CCL5 和 CXCL10 的 mRNA 表达水平。因此，贺芹素与 TNF-α协同作用通过抑制 HeLa 和 T98G 细胞中的 TAK1/IKK/NF-κB 和 TAK1/EGFR 信号通路促进凋亡，为癌症的治疗提供了一种潜在的策略。
