肿瘤细胞治疗研究
英文原题：Paracetamol and Metformin Reduce NK-Cell Susceptibility in MCF-7 Breast Cancer Cells in Association with Enrichment of Immune-Evasive CD44(+)CD24(-) Stem-like Subpopulations.
Paracetamol and Metformin Reduce NK-Cell Susceptibility in MCF-7 Breast Cancer Cells in Association with Enrichment of Immune-Evasive CD44(+)CD24(-) Stem-like Subpopulations.
APAP和MET分别显著富集CD44+CD24- CSC样亚群至75.31%和68.31%，而对照组为11.00%。
自然杀伤（NK）细胞介导的免疫监视是抗肿瘤防御的基石，然而其效力可能受到常见临床药物的损害。对乙酰氨基酚（APAP）和二甲双胍（MET）广泛用于癌症患者的疼痛和代谢管理，但其对免疫-肿瘤界面的非预期效应仍知之甚少。MCF-7乳腺癌细胞（Luminal A亚型）经APAP或MET处理后，与扩增的原代NK细胞（CD3 - CD56 + CD16 +）共培养。我们评估了细胞增殖、细胞周期分布以及CD44 + CD24 - 癌症干细胞样细胞（CSC样）亚群的富集情况。通过RT-qPCR定量了免疫检查点（PD-L1/L2）、应激配体（MICA/B）和共刺激分子CD80/86的转录变化。尽管两种药物均抑制MCF-7生长（48 h IC 50：APAP为11.86 mM；MET为21.11 mM），但二者通过促进免疫逃逸表型诱导了“治疗悖论”。APAP和MET分别将CD44 + CD24 - CSC样亚群显著富集至75.31%和68.31%，而对照组为11.00%。分子分析显示PD-L1（APAP上调19.9倍）和PD-L2（MET上调16.4倍）显著上调，同时CD80/86和MICA/B转录增加。因此，药物处理后的细胞对NK介导的凋亡和坏死表现出显著抵抗。NK细胞优先清除非干细胞（非CSC），无意中进一步富集了高度抵抗的CSC样亚群。额外实验揭示，APAP直接损害NK细胞存活并降低CD3 - CD56 + 细胞比例，而MET对NK细胞影响极小，提示所观察到的NK介导细胞毒性降低存在不同机制。在本研究所采用的实验条件下，APAP 和 MET 通过不同但部分重叠的机制与 MCF-7 细胞对 NK 介导杀伤的易感性降低相关，这些机制包括 CSC 样富集和免疫逃逸通路的转录激活。尽管这些发现是在使用超生理药物浓度的体外机制模型中获得的，但它们确定了潜在机制，值得在生理相关实验系统和体内模型中进一步验证。
Natural Killer (NK) cell-mediated immunosurveillance is a cornerstone of anti-tumor defense, yet its efficacy can be compromised by common clinical medications. Paracetamol (APAP) and metformin (MET) are widely used for pain and metabolic management in cancer patients, but their unintended effects on the immune-tumor interface remain poorly understood. MCF-7 breast cancer cells (Luminal A subtype) were treated with APAP or MET and co-cultured with primary expanded NK cells (CD3 - CD56 + CD16 + ). We evaluated cell proliferation, cell cycle distribution, and the enrichment of the CD44 + CD24 - cancer stem-like cell (CSC-like) subpopulation. Transcriptional changes in immune checkpoints ( PD-L1/L2 ), stress ligands ( MICA/B ), and costimulatory molecules CD80/86 were quantified via RT-qPCR. Despite inhibiting MCF-7 growth (IC 50 at 48 h: 11.86 mM for APAP; 21.11 mM for MET), both drugs induced a "therapeutic paradox" by promoting an immune-evasive phenotype. APAP and MET significantly enriched the CD44 + CD24 - CSC-like subpopulation to 75.31% and 68.31%, respectively, compared to 11.00% in controls. Molecular analysis revealed a robust upregulation of PD-L1 (19.9-fold by APAP) and PD-L2 (16.4-fold by MET), alongside increased CD80/86 and MICA/B transcription. Consequently, drug-treated cells exhibited marked resistance to NK-mediated apoptosis and necrosis. NK cells preferentially eliminated non-stem cells (non-CSCs), inadvertently further concentrating the highly resistant CSC-like subpopulation. Additional experiments revealed that APAP directly impaired NK-cell survival and reduced the proportion of CD3 - CD56 + cells, whereas MET exerted minimal effects on NK cells, suggesting distinct mechanisms underlying the observed reduction in NK-mediated cytotoxicity. Under the experimental conditions employed in this study, APAP and MET were associated with reduced susceptibility of MCF-7 cells to NK-mediated killing through distinct but partially overlapping mechanisms, including CSC-like enrichment and transcriptional activation of immune-evasion pathways. Although these findings were obtained in a mechanistic in vitro model using supra-physiological drug concentrations, they identify potential mechanisms that warrant further validation in physiologically relevant experimental systems and in vivo models.
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