黑色素瘤是最致命的皮肤癌之一，有可能在黑色素细胞存在的任何区域发生。目前，由于全身耐药性的出现而导致的术后复发是黑色素瘤治疗的重大挑战。在这项研究中，从天然对三联苯代谢物中鉴定出了对黑色素瘤细胞具有独特抑制作用的三联苯 (TER)。本研究旨在阐明这种抑制作用的内在机制，这可能有助于发现新型化疗药物。对 TER 处理的 A375 细胞进行转录组测序和代谢组学分析，以确定潜在的作用途径。使用 CRISPR-Cas9 技术和分子克隆敲除并回填关键蛋白质。随后，采用胞质活力、LDH 释放、免疫荧光和流式细胞术的结果来证明药物处理细胞的细胞死亡状态。TER 处理后 p53 信号通路显着上调，导致 CASP3 通过内在的激活凋亡途径。激活的 CASP3 引发细胞凋亡，同时继续裂解 GSDME，从而引发细胞焦亡。 p53（位于该通路上游的关键蛋白）的敲除可显着挽救 TER 诱导的细胞死亡，并减轻细胞活力的下降。然而，位于该通路下游的关键蛋白（CASP3 和 GSDME）的敲除并没有导致 TER 诱导的细胞死亡的挽救，而是导致细胞从凋亡和焦亡转变。 A375 中凋亡和焦亡的诱导TER 对细胞的作用是通过 p53-BAX/FAS-CASP3-GSDME 信号通路介导的。这为TER未来作为潜在的抗黑色素瘤药物奠定了基础。需要注意的是，该通路中的CASP3和GSDME仅调节细胞死亡的方式，而不决定细胞死亡是否发生。这种区别可能在未来的细胞凋亡和焦亡研究中有价值。© 2024。作者。

Melanoma, one of the most lethal forms of skin cancer, has the potential to develop in any area where melanocytes are present. Currently, postoperative recurrence due to the emergence of systemic drug resistance represents a significant challenge in the treatment of melanoma. In this study, terphenyllin (TER), a distinctive inhibitory impact on melanoma cells was identified from the natural p-terphenyl metabolite. This study aimed to elucidate the intrinsic mechanism of this inhibitory effect, which may facilitate the discovery of novel chemotherapeutic agents.A transcriptome sequencing and metabolomic analysis of TER-treated A375 cells was conducted to identify potential pathways of action. The key proteins were knocked out and backfilled using CRISPR-Cas9 technology and molecular cloning. Subsequently, the results of cytosolic viability, LDH release, immunofluorescence and flow cytometry were employed to demonstrate the cell death status of the drug-treated cells.The p53 signalling pathway was markedly upregulated following TER treatment, leading to the activation of CASP3 via the intrinsic apoptotic pathway. The activated CASP3 initiated apoptosis, while simultaneously continuing to cleave the GSDME, thereby triggering pyroptosis. The knockout of p53, a key protein situated upstream of this pathway, resulted in a significant rescue of TER-induced cell death, as well as an alleviation of the decrease in cell viability. However, the knockout of key proteins situated downstream of the pathway (CASP3 and GSDME) did not result in a rescue of TER-induced cell death, but rather a transformation of the cells from apoptosis and pyroptosis.The induction of apoptosis and pyroptosis in A375 cells by TER is mediated via the p53-BAX/FAS-CASP3-GSDME signalling pathway. This lays the foundation for TER as a potential anti-melanoma drug in the future. It should be noted that CASP3 and GSDME in this pathway solely regulate the mode of cell death, rather than determine whether cell death occurs. This distinction may prove valuable in future studies of apoptosis and pyroptosis.© 2024. The Author(s).