银纳米颗粒（AgNPs）通过胞饮作用被小窝依赖内吞作用摄取，这是它们对去势抵抗性前列腺癌具有选择性作用的原因。
Silver Nanoparticles (AgNPs) Uptake by Caveolae-Dependent Endocytosis is Responsible for Their Selective Effect Towards Castration Resistant Prostate Cancer.
机构信息
Molecular Oncology and Viral Pathology Group, Research Center of IPO Porto (CI-IPOP) / RISE@CI-IPOP (Health Research Network), Portuguese Oncology Institute of Porto (IPO Porto) / Porto Comprehensive Cancer Center (Porto.ccc), Porto, Portugal.
ICBAS, Abel Salazar Institute for the Biomedical Sciences, University of Porto, Porto, Portugal.
出版信息
Int J Nanomedicine. 2024 Sep 5;19:9091-9107. doi: 10.2147/IJN.S447645. eCollection 2024.
PURPOSE
Castration Resistant Prostate Cancer (CRPC) is characterized by poor prognosis and limited therapeutic options. AgNPs functionalized with glucose (G-AgNPs) were observed cytotoxic to CRPC cell lines (PC-3 and Du-145) and not LNCaP. This study aims to evaluate AgNPs and G-AgNPs' uptake mechanisms in these cells and understand their role in the selective effect against CRPC cells.
METHODS
Uptake of AgNPs and G-AgNPs was assessed through transmission electron microscopy (TEM). A microRNA (miRNAs) analysis approach was used to uncover the main molecular differences responsible for the endocytic mechanisms' regulation. Caveolin (Cav) 1 and 2 mRNA and protein levels were assessed in the three cell lines. Caveolae-dependent endocytosis was inhibited with genistein or siCav1 and siCav2 in PC-3 and Du-145 and resazurin assay was used to evaluate viability after AgNPs and G-AgNPs administration. Caveolae-dependent endocytosis was induced with Cav1 and Cav2 plasmids in LNCaP, resazurin assay was used to evaluate viability after AgNPs and G-AgNPs administration and TEM to assess their location.
RESULTS
AgNPs and G-AgNPs were not uptaked by LNCaP. miRNA analysis revealed 37 upregulated and 90 downregulated miRNAs. Functional enrichment analysis of miRNAs' targets resulted in enrichment of terms related to endocytosis and caveolae. We observed that Cav1 and Cav2 are not expressed in LNCaP. Inhibiting caveolae-dependent endocytosis in Du-145 and PC-3 led to a significative reduction of cytotoxic capacity of AgNPs and G-AgNPs and induction of caveolae-dependent endocytosis in LNCaP lead to a significative increase as well as their uptake by cells.
CONCLUSION
This study shows the potential of these AgNPs as a new therapeutic approach directed to CRPC patients, uncovers caveolae-dependent endocytosis as the uptake mechanism of these AgNPs and highlights deregulation of Cav1 and Cav2 expression as a key difference in hormone sensitive and resistant PCa cells which may be responsible for drug resistance.
目的
去势抵抗性前列腺癌（CRPC）的预后较差，治疗选择有限。研究发现，葡萄糖功能化的银纳米颗粒（G-AgNPs）对 CRPC 细胞系（PC-3 和 Du-145）具有细胞毒性，而对 LNCaP 则没有。本研究旨在评估 AgNPs 和 G-AgNPs 在这些细胞中的摄取机制，并了解其在对 CRPC 细胞的选择性作用中的作用。
方法
通过透射电子显微镜（TEM）评估 AgNPs 和 G-AgNPs 的摄取。采用 microRNA（miRNAs）分析方法揭示了主要的分子差异，这些差异负责调节内吞作用的机制。评估了三种细胞系中 Cavlin1 和 2 的 mRNA 和蛋白水平。用染料木黄酮或 siCav1 和 siCav2 抑制 PC-3 和 Du-145 中的 caveolae 依赖性内吞作用，并使用 Resazurin 测定法评估 AgNPs 和 G-AgNPs 给药后的细胞活力。用 Cav1 和 Cav2 质粒诱导 LNCaP 中的 caveolae 依赖性内吞作用，用 Resazurin 测定法评估 AgNPs 和 G-AgNPs 给药后的细胞活力，并通过 TEM 评估其位置。
结果
AgNPs 和 G-AgNPs 不能被 LNCaP 摄取。miRNAs 分析显示 37 个上调和 90 个下调 miRNAs。miRNAs 靶标的功能富集分析导致与内吞作用和 caveolae 相关的术语富集。我们观察到 Cavlin1 和 Cavlin2 不在 LNCaP 中表达。在 Du-145 和 PC-3 中抑制 caveolae 依赖性内吞作用会导致 AgNPs 和 G-AgNPs 的细胞毒性显著降低，而在 LNCaP 中诱导 caveolae 依赖性内吞作用会导致细胞毒性显著增加以及细胞摄取 AgNPs 和 G-AgNPs 的增加。
结论
本研究表明，这些 AgNPs 具有作为治疗 CRPC 患者的新方法的潜力，揭示了 caveolae 依赖性内吞作用是这些 AgNPs 的摄取机制，并强调了 Cavlin1 和 Cavlin2 表达的下调是激素敏感和耐药 PCa 细胞的关键差异，这可能是耐药的原因。
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