Hao Yuanyuan, Duan Feng, Dong Xianning, Bi Ran, Wang Yinzhe, Zhu Senqiang, Hu Jinghai
Department of Urology, The First Hospital of Jilin University, Changchun, China.
Department of Oncology, Qingdao Municipal Hospital, Qingdao, China.
Anal Cell Pathol (Amst). 2025 Jan 4;2025:6648632. doi: 10.1155/ancp/6648632. eCollection 2025.
This study aims to study how gold nanoparticles (AuNPs) function in the recruitment and polarization of tumor-associated macrophages (TAMs) in hormone-sensitive prostate cancer (HSPC) and castration-resistant prostate cancer (CRPC). Phorbol ester (PMA)-treated THP-1 cells were cocultured with LNCaP or PC3 cells to simulate TAMs. Macrophage M2 polarization levels were detected using flow cytometry and M2 marker determination. ATG5 expression was detected by western blotting. Luciferase reporter assay was used to analyze the N6-methyladenosine (mA) site activity of ATG5 3' untranslated regions (3'-UTRs). Methylated RNA immune precipitation (MeRIP)-quantitative polymerase chain reaction (qPCR) was performed to determine the mA levels at ATG5 3'-UTR. Xenograft mouse models were used to determine the function of AuNPs in vivo. Macrophages exhibited reduced M2 polarization in both HSPC and CRPC cells after AuNP treatment which was prevented by induction of autophagy. AuNP treatment decreased the mA levels in the 3'-UTR of ATG5. Mutational analysis of potential mA sites within ATG5 3'-UTR revealed that these sites were required for AuNP regulation, indicating that AuNPs inhibited ATG5 levels in an mA-dependent manner. The mouse model revealed that AuNPs significantly reduced the M2 polarization of TAMs in an autophagy-dependent manner in vivo. This suggests that AuNPs inhibit tumor growth in vivo partially through targeting M2 TAM. The ATG5/autophagy pathway is inhibited by AuNP treatment in an METTL3/mA-dependent manner. AuNPs inhibit the TAM M2 polarization in HSPC and CRPC by inhibiting ATG5/autophagy.
本研究旨在探讨金纳米颗粒（AuNPs）在激素敏感性前列腺癌（HSPC）和去势抵抗性前列腺癌（CRPC）中对肿瘤相关巨噬细胞（TAMs）的募集和极化作用。用佛波酯（PMA）处理的THP-1细胞与LNCaP或PC3细胞共培养以模拟TAMs。采用流式细胞术和M2标志物测定法检测巨噬细胞M2极化水平。通过蛋白质免疫印迹法检测自噬相关蛋白5（ATG5）的表达。使用荧光素酶报告基因检测法分析ATG5 3'非翻译区（3'-UTR）的N6-甲基腺苷（m⁶A）位点活性。进行甲基化RNA免疫沉淀（MeRIP）-定量聚合酶链反应（qPCR）以测定ATG5 3'-UTR处的m⁶A水平。利用异种移植小鼠模型确定AuNPs在体内的功能。AuNP处理后，HSPC和CRPC细胞中的巨噬细胞M2极化均降低，而自噬诱导可阻止这种降低。AuNP处理降低了ATG5 3'-UTR中的m⁶A水平。对ATG5 3'-UTR内潜在m⁶A位点的突变分析表明，这些位点是AuNP调控所必需的，这表明AuNPs以m⁶A依赖的方式抑制ATG5水平。小鼠模型显示，AuNPs在体内以自噬依赖的方式显著降低了TAMs的M2极化。这表明AuNPs在体内部分通过靶向M2型TAM抑制肿瘤生长。AuNP处理以甲基转移酶样3（METTL3）/m⁶A依赖的方式抑制ATG5/自噬途径。AuNPs通过抑制ATG5/自噬来抑制HSPC和CRPC中的TAM M2极化。
