RNA-Seq 探索基于一体化纳米气泡的氧增强声动力学疗法增强铁死亡治疗 HCC 的机制。

BACKGROUND: The combination of sonodynamic therapy and oxygenation strategy is widely used in cancer treatment. However, due to the complexity, heterogeneity and irreversible hypoxic environment produced by hepatocellular carcinoma (HCC) tissues, oxygen-enhancing sonodynamic therapy (SDT) has failed to achieve the desired results. With the emergence of ferroptosis with reactive oxygen species (ROS) cytotoxicity, this novel cell death method has attracted widespread attention. METHODS: In this study, nanobubbles (NBs) were connected with the sonosensitizer Indocyanine green (ICG) to construct a 2-in-1 nanoplatform loaded with RAS-selective lethal (RSL3, ferroptosis promoter) (RSL3@O2-ICG NBs), combined with oxygen-enhanced SDT and potent ferroptosis. In addition, nanobubbles (NBs) combined with low-frequency ultrasound (LFUS) are called ultrasound-targeted nanobubble destruction (UTND) to ensure specific drug release and improve safety. RESULTS: MDA/GSH and other related experimental results show that RSL3@O2-ICG NBs can enhance SDT and ferroptosis. Through RNA sequencing (RNA-seq), the differential expression of LncRNA and mRNA before and after synergistic treatment was identified, and then GO and KEGG pathways were used to enrich and analyze target genes and pathways related ferroptosis sensitivity. We found that they were significantly enriched in the ferroptosis-related pathway MAPK cascade and cell proliferation. Then, we searched for the expression of differentially expressed genes in the TCGA Hepatocellular carcinoma cohort. At the same time, we evaluated the proportion of immune cell infiltration and the identification of co-expression network modules and related prognostic analysis. We found that it was significantly related to the tumor microenvironment of hepatocellular carcinoma. The prognostic risk genes "SLC37A2" and "ITGB7" may represent new hepatocellular carcinoma ferroptosis-inducing markers and have guiding significance for treating hepatocellular carcinoma. CONCLUSION: The therapeutic effect of the in vitro synergistic treatment has been proven to be significant, revealing the prospect of 2-in-1 nanobubbles combined with SDT and ferroptosis in treating HCC.

背景：声动力学疗法与氧合策略的联合应用广泛应用于癌症治疗中。然而，由于肝癌（HCC）组织的复杂性、异质性和不可逆转的缺氧环境，增强氧的声动力学疗法（SDT）未能达到预期的效果。随着具有活性氧（ROS）细胞毒性的铁死亡的出现，这种新的细胞死亡方法引起了广泛的关注。

方法：在这项研究中，纳米气泡（NBs）与声敏剂吲哚菁绿（ICG）相连，构建了一种负载 RAS 选择性致死（RSL3，铁死亡促进剂）（RSL3@O2-ICG NBs）的 2-in-1 纳米平台，结合增强氧的 SDT 和强大的铁死亡。此外，纳米气泡（NBs）与低频超声（LFUS）结合称为超声靶向纳米气泡破坏（UTND），以确保特定药物的释放并提高安全性。

结果：MDA/GSH 等相关实验结果表明，RSL3@O2-ICG NBs 可增强 SDT 和铁死亡。通过 RNA 测序（RNA-seq），鉴定了协同治疗前后 LncRNA 和 mRNA 的差异表达，然后使用 GO 和 KEGG 途径对与铁死亡敏感性相关的靶基因和途径进行富集和分析。我们发现它们在铁死亡相关途径 MAPK 级联和细胞增殖中显著富集。然后，我们在 TCGA 肝癌队列中搜索差异表达基因的表达。同时，我们评估了免疫细胞浸润的比例，识别共表达网络模块并进行相关的预后分析。我们发现它与肝癌的肿瘤微环境显著相关。预后风险基因“ SLC37A2”和“ ITGB7”可能代表新的肝癌铁死亡诱导标志物，对治疗肝癌具有指导意义。

结论：体外协同治疗的疗效已被证明是显著的，揭示了 2-in-1 纳米气泡与 SDT 和铁死亡联合治疗 HCC 的前景。