PO.PR02.01 · 预防研究
水飞蓟宾诱导的肥大细胞调控：在胰腺癌预防与治疗中经ODC1实现潜在免疫调节
Silibinin induced mast cell regulation: Potential immunomodulation via ODC1 in pancreatic cancer prevention and therapy
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
美国癌症协会估计，2025年美国将有约67,440人（34,950名男性；32,490名女性）被诊断为胰腺癌（PC），约51,980人（27,050名男性；24,930名女性）将死于胰腺癌。此外，据估计发生PC的平均终生风险在男性中约为1/56，在女性中约为1/60。据报道，PC是美国癌症死亡的第4大原因，总体5年生存率<5%。免疫系统，尤其是初级细胞反应者肥大细胞（MC），在PC中的作用仍不明确。天然黄酮木脂素水飞蓟宾（SB）在PC的临床前模型中有效；然而，据我们所知，其在PC免疫调节中的作用尚未见报道，尤其是关于MC的作用。我们此前报道了SB在基底细胞癌、前列腺癌、结肠癌和肝细胞癌中的免疫调节作用。因此，我们利用源自C57BL/6小鼠的骨髓肥大细胞（BMMC），在IL-3和SCF存在下（通过流式细胞术[cKit/FcεRI双染]确认），确定了SB处理对PC中MC的机制层面。BMMC暴露于不同剂量的SB（25-100 μM），并选择两种浓度（25 μM，SB 25和100 μM，SB 100）进行进一步实验。将经SB 25、SB 100处理及未处理（对照）的BMMC进行蛋白质组学分析（在Fusion Lumos质谱仪上进行LCMS）；使用注释的小鼠蛋白质组鉴定出3575个蛋白质。统计分析（ANOVA后接Fisher事后分析；p<0.05，FDR<0.01）发现166个蛋白质在三组间存在统计学显著的差异表达（n=4/组）。选择这166个蛋白质使用Ingenuity Pathway Analysis（IPA；Qiagen）进行进一步的通路分析。仅选择在PC免疫细胞/细胞系中经实验证实的通路进行进一步分析。发现与SB处理经MC进行免疫细胞调控相关的主要经典通路为叶酸信号通路、衰老相关分泌表型、细胞周期蛋白与细胞周期调控、RUNX2的转录调控以及衰老通路。发现重要的上游调控因子为ODC1、CDKN1A、SMAD2、PLAC8、TP53。确定的重要节点为KRAS、GLI1和ODC1，所涉及的主要因果网络为多胺合成，其枢纽为鸟氨酸脱羧酶1（ODC1）。ODC1抑制被提出为一种潜在的治疗策略，因为胰腺肿瘤，尤其是那些对常规治疗耐药的肿瘤，高度依赖此酶实现肿瘤进展。因此，SB可能通过调控MC和ODC1在PC中发挥重要的免疫调节作用。有必要开展进一步研究以阐明这些机制，将SB作为一种预防或治疗选择加以利用（由R01 CA140368资助）。
查看英文原文 English abstract
It has been estimated by the American Cancer Society that in the US in 2025, ~67,440 people (34,950 men; 32,490 women) will be diagnosed with, and ~51,980 people (27,050 men; 24,930 women) will die of pancreatic cancer (PC). Additionally, the average lifetime risk of developing PC has been estimated to be ~1 in 56 in men and ~1 in 60 in women. PC has been reported to be the 4th leading cause of cancer deaths in the US with a <5% overall 5-year survival rate. The role of immune system, particularly the primary cellular responders, the mast cells (MCs), in PC remains obscure. The natural flavonolignan silibinin (SB), has been effective in preclinical models of PC; however, its role in the immune modulation in PC has not been reported, to our knowledge, particularly with respect to MCs. We previously reported immunomodulation by SB in basal cell carcinoma, prostate cancer, colon cancer, and hepatocellular carcinoma. Thus, mechanistic aspects of SB treatment on MCs in PC were determined, leveraging bone marrow MCs (BMMCs) derived from C57BL/6 mice, in the presence of IL-3 and SCF (confirmed using flow cytometry [cKit/Fc€RI dual staining]). BMMCs were exposed to different doses of SB (25-100 µM) and two concentrations (25 µM; SB 25 and 100 µM; SB 100) were selected for further experiments. BMMCs treated with SB 25, SB 100, and untreated (control) were subjected to proteomics (LCMS on Fusion Lumos mass spectrometer); 3575 proteins were identified using the annotated mouse proteome. Statistical analysis (ANOVA followed by Fisher's posthoc analysis; p<0.05, FDR<0.01) and 166 proteins were found to have statistically significant differential expressions amongst the three groups (n=4/group). These 166 proteins were selected for further pathway analysis using Ingenuity Pathway Analysis (IPA; Qiagen). Only pathways specifically demonstrated experimentally in immune cells/cell lines in PC were selected for further analysis. Top canonical pathways associated with SB treatment of immune cell regulation via MCs were found to be folate signaling pathway, senescence-associated secretory phenotype, cyclins and cell cycle regulation, transcriptional regulation by RUNX2, as senescence pathway. Important upstream regulators were found to be ODC1, CDKN1A, SMAD2, PLAC8, TP53. Important nodes determined were KRAS, GLI1, and ODC1, with the main causal network implicated being polyamine synthesis, hinged on ornithine decarboxylase 1 (ODC1). ODC1 inhibition is proposed to be a potential therapeutic strategy, as pancreatic tumors particularly those resistant to conventional treatments are highly dependent on this enzyme for tumor progression. Thus, SB may play an important immunomodulatory role in PC via regulation of MCs and ODC1. Further studies are warranted to delineate these mechanisms leveraging SB in the form of a preventive or treatment option (supported by R01 CA140368).
利益披露 Disclosure
N. Mishra, None..
S. Paudel, None..
C. Agarwal, None..
R. Agarwal, None.