AKR1C3酶在激素和药物代谢中起着至关重要的作用,并与肝癌的异常表达有关,导致肿瘤进展和预后不良。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和减轻缺氧。因此,通过构建HSA-RSVNPs载体,探索白藜芦醇对AKR1C3的潜在调控机制,对肝癌的治疗具有重要的理论和临床意义。这项研究的目的是研究通过负载白藜芦醇(RSV)在纳米材料HSA-RSVNPs(纳米颗粒)上靶向调节AKR1C3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),从GeneCards数据库中检索到相关基因,导致三个相交以获得候选DEG。对候选DEGs进行GO和KEGG富集分析,以分析受主要靶基因影响的潜在细胞功能和分子信号通路。使用cytohubba插件筛选按Degree排序的前10个目标基因,并进一步交叉LASSO和随机森林(RF)的结果以获得hub基因。进行hub基因的表达分析及对恶性肿瘤预后的预测。此外,使用PharmMapper构建药效团模型。使用AutoDockTools1.5.6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细胞系进行体外细胞实验,用不同浓度的RSV治疗HCC细胞,或使用慢病毒沉默或过表达AKR1C3。CCK-8,克隆形成,流式细胞术,划痕实验,Transwell被用来测量癌细胞的活力,扩散,迁移,入侵,和细胞凋亡,分别。使用海马XF24分析仪分析细胞耗氧率。制备HSA-RSVNP,并对其表征和细胞毒性进行了评估。检测治疗后肝癌细胞的生物学功能变化。使用HepG2细胞系在小鼠中建立HCC皮下异种移植模型。经尾静脉注射HSA-RSVNP,使用控制组集,观察肿瘤生长的变化,NPs的肿瘤靶向,生物安全。TUNEL,在切除的肿瘤组织上进行Ki67和APC-缺氧探针染色以检测肿瘤细胞的增殖。凋亡,和缺氧。慢病毒用于沉默或过表达AKR1C3,同时通过尾静脉注射HSA-RSVNP,以评估AKR1C3对HCC进展中HSA-RSVNP调节的影响。生物信息学分析显示,AKR1C3是RSV调控HCC的重要靶基因,这与HCC患者的预后有关,并且表达上调。体外细胞实验表明,RSV显著抑制HCC细胞的呼吸代谢,抑制它们的扩散,迁移,侵袭和促进细胞凋亡。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证明了HSA-RSVNP的成功构建,对肝癌细胞有较强的抑制作用。在体内,动物实验进一步证实,HSA-RSVNP靶向下调AKR1C3抑制HCC进展和肿瘤缺氧,同时表现出肿瘤靶向性和生物学安全性。HSA-RSVNPs靶向下调AKR1C3可减轻HCC肿瘤缺氧并抑制HCC进展。
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.