Hepatocellular carcinoma (HCC) is a severe disease that accounts for 80% of liver cancers. Chemotherapy is the primary therapeutic strategy for patients who cannot be treated with surgery or who have late-stage HCC. C₂-ceramide is an effective reagent that has been found to inhibit the growth of many cancer types. The metabolism of C₂-ceramide plays a vital role in the regulation of cell death/cell survival. The phenoxyphenol compound 4-{2,3,5,6-tetrafluoro-4-[2,3,5,6-tetrafluoro-4-(4-hydroxyphenoxy)phenyl]phenoxy}phenol (diTFPP) was found to have a synergistic effect with C₂-ceramide, resulting in considerable cell death in the HA22T HCC cell line. diTFPP/C₂-ceramide cotreatment induced a two- to threefold increase in cell death compared to that with C2-ceramide alone and induced pyknosis. Annexin V/7-aminoactinomycin D (7AAD) double staining and Western blotting indicated that apoptosis was involved in diTFPP/C₂-ceramide cotreatment-mediated cell death. We next analyzed transcriptome alterations in diTFPP/C₂-ceramide-cotreated HA22T cells with next-generation sequencing (NGS). The data indicated that diTFPP treatment disrupted sphingolipid metabolism, inhibited cell cycle-associated gene expression, and induced autophagy and reactive oxygen species (ROS)-responsive changes in gene expression. Additionally, we assessed the activation of autophagy with acridine orange (AO) staining and observed alterations in the expression of the autophagic proteins LC3B-II and Beclin-1, which indicated autophagy activation after diTFPP/C₂-ceramide cotreatment. Elevated levels of ROS were also reported in diTFPP/C₂-ceramide-treated cells, and the expression of the ROS-associated proteins SOD1, SOD2, and catalase was upregulated after diTFPP/C₂-ceramide treatment. This study revealed the potential regulatory mechanism of the novel compound diTFPP in sphingolipid metabolism by showing that it disrupts ceramide metabolism and apoptotic sphingolipid accumulation.

中文翻译：

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苯氧酚化合物diTFPP介导肝细胞癌细胞外源性C2-神经酰胺代谢，并伴随着ROS的形成和自噬而诱导细胞凋亡。

肝细胞癌（HCC）是一种严重疾病，占肝癌的80％。对于不能接受手术治疗或晚期肝癌的患者，化学疗法是主要的治疗策略。C _2-神经酰胺是一种有效的试剂，已发现可以抑制许多癌症类型的生长。C _2-神经酰胺的代谢在调节细胞死亡/细胞存活中起着至关重要的作用。发现苯氧基苯酚化合物4- {2,3,5,6-四氟-4- [2,3,5,6-四氟-4-（4-羟基苯氧基）苯基]苯氧基}苯酚（diTFPP）具有协同作用C _2-神经酰胺的作用，导致HA22T HCC细胞系中大量细胞死亡。diTFPP / C ₂与单独使用C2-神经酰胺的情况相比，神经酰胺共处理诱导的细胞死亡增加了2到3倍，并引起了k缩。Annexin V / 7-氨基放线菌素D（7AAD）双重染色和Western印迹表明凋亡与diTFPP / C _2-神经酰胺共同治疗介导的细胞死亡有关。接下来，我们分析了diTFPP / C _2中的转录组变化-神经酰胺处理的HA22T细胞具有下一代测序（NGS）功能。数据表明，diTFPP处理破坏了鞘脂代谢，抑制了与细胞周期相关的基因表达，并诱导了基因表达中的自噬和活性氧（ROS）响应变化。此外，我们用with啶橙（AO）染色评估了自噬的激活，并观察到自噬蛋白LC3B-II和Beclin-1的表达发生了变化，这表明diTFPP / C _2-神经酰胺共处理后自噬的激活。ROS水平升高也报告在diTFPP / C ₂ -神经酰胺处理的细胞，并且ROS相关蛋白SOD1，SOD2，和过氧化氢的表达diTFPP / C后上调₂-神经酰胺治疗。这项研究揭示了新型化合物diTFPP破坏神经酰胺代谢和凋亡性鞘脂蓄积，从而揭示了其在鞘脂代谢中的潜在调控机制。