本研究旨在开发一种高效的肿瘤靶向脂质体纳米气泡 (LNB) 系统，利用超声靶向纳米气泡破坏来增强肝细胞癌 (HCC) 治疗中 miRNA-199a-3p 的释放和转染。制备的 LNB 包含聚乙二醇修饰的脂质体壳和全氟戊烷（PFP）核。 miRNA-199a-3p 通过静电吸附附着在纳米复合材料表面，而 RGD 肽功能化 LNB 表面以增强 HCC 细胞靶向，即 PFP@miR-RGD-LNBs。 LNB 呈球形，尺寸分布较窄。负载基因的 LNB 有效地浓缩了 miR-199a-3p 并保护其免受酶降解。低强度聚焦超声（LIFU）促进miR-199a-3p从制备的LNB中快速释放，从而增强治疗效果。 PFP@miR-RGD-LNBs 和 LIFU 的联合应用对 HepG2 细胞表现出比其他组更有效的抑制作用，这可能是由于 LIFU 促进靶位点快速有效的基因释放并增加细胞膜通透性。定量逆转录聚合酶链反应分析显示，与其他组相比，PFP@miR-RGD-LNBs  LIFU 组中关键凋亡标志物（Bad、Bax、Caspase-9 和 Caspase-3）的 mRNA 表达水平显着增加。这些发现表明，制备的 LNB 很可能成为进一步探索 HCC 基因传递和治疗的有希望的候选者。

This study was aimed to develop an efficient tumour-targeted liposome nanobubbles (LNBs) system using ultrasound-targeted nanobubble destruction for enhanced release and transfection of miRNA-199a-3p in hepatocellular carcinoma (HCC) therapy. The prepared LNBs comprised a polyethylene glycol-modified liposome shell and a perfluoropentane (PFP) core. MiRNA-199a-3p was attached to the nanocomposite surface via electrostatic adsorption, while RGD peptide functionalized the LNBs surface for enhanced HCC cell targeting, namely PFP@miR-RGD-LNBs. The LNBs were spherical with a narrow size distribution. The gene-loaded LNBs effectively condensed miR-199a-3p and protected it from enzymatic degradation. Low-intensity focused ultrasound (LIFU) promoted a fast release of miR-199a-3p from the prepared LNBs, thereby enhancing therapeutic effects. The combined application of PFP@miR-RGD-LNBs and LIFU exhibited a more potent inhibitory effect on HepG2 cells than the other groups, potentially due to LIFU promoting rapid and efficient gene release at the target site and increasing cell membrane permeability. Quantitative reverse transcription-polymerase chain reaction analysis revealed significantly increased mRNA expression levels of key apoptosis markers (Bad, Bax, Caspase-9 and Caspase-3) in the PFP@miR-RGD-LNBs + LIFU group compared to other groups. These findings suggest that the prepared LNBs are highly likely to be promising candidates for further exploration of HCC gene delivery and therapy.