Operating Room, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, 121001, China.
Department of Critical Care Medicine, Peking Union Medical College Hospital, Beijing, 100730, China.
Apoptosis. 2024 Oct;29(9-10):1429-1453. doi: 10.1007/s10495-024-01995-w. Epub 2024 Jul 18.
The enzyme AKR1C3 plays a crucial role in hormone and drug metabolism and is associated with abnormal expression in liver cancer, leading to tumor progression and poor prognosis. Nanoparticles modified with HSA can modulate the tumor microenvironment by enhancing photodynamic therapy to induce apoptosis in tumor cells and alleviate hypoxia. Therefore, exploring the potential regulatory mechanisms of resveratrol on AKR1C3 through the construction of HSA-RSV NPs carriers holds significant theoretical and clinical implications for the treatment of liver cancer. The aim of this study is to investigate the targeted regulation of AKR1C3 expression through the loading of resveratrol (RSV) on nanomaterials HSA-RSV NPs (Nanoparticles) in order to alleviate tumor hypoxia and inhibit the progression of hepatocellular carcinoma (HCC), and to explore its molecular mechanism. PubChem database and PharmMapper server were used to screen the target genes of RSV. HCC-related differentially expressed genes (DEGs) were analyzed through the GEO dataset, and relevant genes were retrieved from the GeneCards database, resulting in the intersection of the three to obtain candidate DEGs. GO and KEGG enrichment analyses were performed on the candidate DEGs to analyze the potential cellular functions and molecular signaling pathways affected by the main target genes. The cytohubba plugin was used to screen the top 10 target genes ranked by Degree and further intersected the results of LASSO and Random Forest (RF) to obtain hub genes. The expression analysis of hub genes and the prediction of malignant tumor prognosis were conducted. Furthermore, a pharmacophore model was constructed using PharmMapper. Molecular docking simulations were performed using AutoDockTools 1.5.6 software, and ROC curve analysis was performed to determine the core target. In vitro cell experiments were carried out by selecting appropriate HCC cell lines, treating HCC cells with different concentrations of RSV, or silencing or overexpressing AKR1C3 using lentivirus. CCK-8, clone formation, flow cytometry, scratch experiment, and Transwell were used to measure cancer cell viability, proliferation, migration, invasion, and apoptosis, respectively. Cellular oxygen consumption rate was analyzed using the Seahorse XF24 analyzer. HSA-RSV NPs were prepared, and their characterization and cytotoxicity were evaluated. The biological functional changes of HCC cells after treatment were detected. An HCC subcutaneous xenograft model was established in mice using HepG2 cell lines. HSA-RSV NPs were injected via the tail vein, with a control group set, to observe changes in tumor growth, tumor targeting of NPs, and biological safety. TUNEL, Ki67, and APC-hypoxia probe staining were performed on excised tumor tissue to detect tumor cell proliferation, apoptosis, and hypoxia. Lentivirus was used to silence or overexpress AKR1C3 simultaneously with the injection of HSA-RSV NPs via the tail vein to assess the impact of AKR1C3 on the regulation of HSA-RSV NPs in HCC progression. Bioinformatics analysis revealed that AKR1C3 is an important target gene involved in the regulation of HCC by RSV, which is associated with the prognosis of HCC patients and upregulated in expression. In vitro cell experiments showed that RSV significantly inhibits the respiratory metabolism of HCC cells, suppressing their proliferation, migration, and invasion and promoting apoptosis. Silencing AKR1C3 further enhances the toxicity of RSV towards HCC cells. The characterization and cytotoxicity experiments of nanomaterials demonstrated the successful construction of HSA-RSV NPs, which exhibited stronger inhibitory effects on HCC cells. In vivo, animal experiments further confirmed that targeted downregulation of AKR1C3 by HSA-RSV NPs suppresses the progression of HCC and tumor hypoxia while exhibiting tumor targeting and biological safety. Targeted downregulation of AKR1C3 by HSA-RSV NPs can alleviate HCC tumor hypoxia and inhibit the progression of HCC.
酶 AKR1C3 在激素和药物代谢中发挥着关键作用,与肝癌中的异常表达有关,导致肿瘤进展和预后不良。通过用 HSA 修饰的纳米粒子,可以通过增强光动力疗法来调节肿瘤微环境,从而诱导肿瘤细胞凋亡并减轻缺氧。因此,通过构建 HSA-RSV NPs 载体来探索白藜芦醇对 AKR1C3 的潜在调节机制,对于治疗肝癌具有重要的理论和临床意义。本研究的目的是通过将白藜芦醇(RSV)载入纳米材料 HSA-RSV NPs(纳米粒子)中来靶向调节 AKR1C3 的表达,以减轻肿瘤缺氧并抑制肝细胞癌(HCC)的进展,并探讨其分子机制。使用 PubChem 数据库和 PharmMapper 服务器筛选 RSV 的靶基因。通过 GEO 数据集分析 HCC 相关差异表达基因(DEGs),并从 GeneCards 数据库中检索相关基因,将三个数据库的结果取交集,得到候选 DEGs。对候选 DEGs 进行 GO 和 KEGG 富集分析,以分析主要靶基因所影响的潜在细胞功能和分子信号通路。使用 cytohubba 插件筛选排名前十的以 Degree 排名的靶基因,并进一步与 LASSO 和随机森林(RF)的结果相交,以获得枢纽基因。分析枢纽基因的表达和恶性肿瘤预后的预测。此外,使用 PharmMapper 构建药效团模型。使用 AutoDockTools 1.5.6 软件进行分子对接模拟,并进行 ROC 曲线分析以确定核心靶标。通过选择合适的 HCC 细胞系,用不同浓度 RSV 处理 HCC 细胞,或使用慢病毒沉默或过表达 AKR1C3,在体外细胞实验中进行。分别用 CCK-8、克隆形成、流式细胞术、划痕实验和 Transwell 实验测量癌细胞活力、增殖、迁移、侵袭和凋亡。使用 Seahorse XF24 分析仪分析细胞耗氧率。制备 HSA-RSV NPs,并对其特性和细胞毒性进行评估。检测 HCC 细胞经处理后的生物学功能变化。使用 HepG2 细胞系建立 HCC 皮下移植瘤模型,通过尾静脉注射 HSA-RSV NPs,并设置对照组,观察肿瘤生长、NPs 肿瘤靶向和生物安全性的变化。对切除的肿瘤组织进行 TUNEL、Ki67 和 APC-缺氧探针染色,以检测肿瘤细胞增殖、凋亡和缺氧。同时使用慢病毒沉默或过表达 AKR1C3,然后通过尾静脉注射 HSA-RSV NPs,评估 AKR1C3 对 HSA-RSV NPs 调节 HCC 进展的影响。生物信息学分析表明,AKR1C3 是 RSV 调节 HCC 的重要靶基因,与 HCC 患者的预后相关,且在表达水平上上调。体外细胞实验表明,RSV 显著抑制 HCC 细胞的呼吸代谢,抑制其增殖、迁移和侵袭,促进凋亡。沉默 AKR1C3 进一步增强 RSV 对 HCC 细胞的毒性。纳米材料的表征和细胞毒性实验表明,成功构建了 HSA-RSV NPs,其对 HCC 细胞具有更强的抑制作用。体内动物实验进一步证实,通过 HSA-RSV NPs 靶向下调 AKR1C3 可抑制 HCC 的进展和肿瘤缺氧,同时具有肿瘤靶向和生物安全性。通过 HSA-RSV NPs 靶向下调 AKR1C3 可减轻 HCC 肿瘤缺氧并抑制 HCC 进展。