METHODS: Cell viability was analyzed by WST-8 assay. Apoptosis induction, BrdU incorporation, and cell cycle analysis were performed using flow cytometry. Metabolites were measured by LC-MS/MS analysis. Real-time PCR and western blotting evaluated RNA and protein expression. DNA damage was quantified through immunofluorescence. cBioPortal and GEPIA data were utilized to explore the mutual co-occurrence of TP53 and UMPS and UMPS gene expression in NSCLC.
RESULTS: The combination treatment suppressed de novo pyrimidine nucleotide biosynthesis by reducing the expression of related enzymes. This blockade of pyrimidine metabolism led to DNA damage and subsequent apoptosis, revealing a novel mechanism for inducing lung cancer cell death with this combination. However, some lung cancer cells exhibited distinct responses to the combination treatment that inhibited pyrimidine metabolism. The differences in sensitivity in lung cancer cells were determined by the TP53 gene status. TP53 wild-type lung cancer cells were effectively inhibited by the combination treatment through p53 activation, while TP53 mutant- or null-type cells exhibited lower sensitivity.
CONCLUSIONS: This study, for the first time, established a link between cancer cell genetic features and treatment response to simultaneous SH003 and docetaxel treatment. It highlights the significance of p53 as a predictive factor for susceptibility to this combination treatment. These findings also suggest that p53 status could serve as a crucial criterion in selecting appropriate therapeutic strategies for targeting pyrimidine metabolism in lung cancer.
方法:通过WST-8测定分析细胞活力。凋亡诱导,BrdU成立,使用流式细胞术进行细胞周期分析。通过LC-MS/MS分析测量代谢物。实时PCR和蛋白质印迹评估RNA和蛋白质表达。通过免疫荧光定量DNA损伤。cBioPortal和GEPIA数据用于探索NSCLC中TP53和UMPS和UMPS基因表达的相互共现。
结果:联合治疗通过减少相关酶的表达来抑制嘧啶核苷酸的从头生物合成。这种嘧啶代谢的阻断导致DNA损伤和随后的细胞凋亡,揭示了这种组合诱导肺癌细胞死亡的新机制。然而,一些肺癌细胞对抑制嘧啶代谢的联合治疗表现出不同的反应。肺癌细胞的敏感性差异由TP53基因状态决定。TP53野生型肺癌细胞通过p53激活的联合治疗被有效抑制,而TP53突变或无效型细胞表现出较低的敏感性。
结论:这项研究,第一次,建立了同时SH003和多西他赛治疗的癌细胞遗传特征和治疗反应之间的联系。它强调了p53作为这种联合治疗易感性的预测因子的重要性。这些发现还表明,p53状态可以作为选择针对肺癌嘧啶代谢的适当治疗策略的关键标准。