The present study aims to develop a nanoparticle encapsulating doxorubicin (DOX) and programmed death-ligand 1 (PD-L1) siRNA and evaluate its anti-tumor effects on hepatoma carcinoma (HCC). Nanoparticle encapsulating DOX and PD-L1 siRNA (NPDOX/siPD-L1) was characterized by dynamic light scattering and transmission electron microscopy. Flow cytometry was applied to analyze cell populations, NPDOX/siPD-L1 internalization, and cell apoptosis. Real-Time (RT)-quantitative reverse transcription (qPCR) and western blotting were used to determine the mRNA and protein levels, respectively. Released ATP was determined using ATP determination kit and cytokines were determined using specific ELISAs. A tumor-bearing animal model was established to evaluate the anti-tumor effects of NPDOX/siPD-L1. Treatment of NPDOX/siPD-L1 induced immunogenic cell death (ICD) and PD-L1 overexpression in HCC. In vivo study demonstrated that intravenously injection of NPDOX/siPD-L1 significantly inhibited the tumor volume and PD-L1 expressions of tumor tissue in the H22 tumor-bearing animal model. Besides, the treatment of NPDOX/siPD-L1 also regulated the populations of matured dendritic cells and cytotoxic T cells and the productions of cytokines in the tumor tissues. Taken together, NPDOX/siPD-L1 showed significant anti-tumor effects on HCC by the induction of ICD and inhibition of PD-L1 overexpression.

中文翻译：

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纳米药物介导的免疫原性细胞死亡诱导和预防PD-L1过表达，以增强肝细胞癌治疗

本研究旨在开发一种封装阿霉素（DOX）和程序性死亡配体1（PD-L1）siRNA的纳米颗粒，并评估其对肝癌（HCC）的抗肿瘤作用。通过动态光散射和透射电子显微镜表征了封装DOX和PD-L1 siRNA（NPDOX / siPD-L1）的纳米颗粒。流式细胞仪用于分析细胞群体，NPDOX / siPD-L1内在化和细胞凋亡。实时（RT）定量逆转录（qPCR）和蛋白质印迹分别用于确定mRNA和蛋白质水平。使用ATP测定试剂盒测定释放的ATP，并使用特异性ELISA测定细胞因子。建立了荷瘤动物模型以评估NPDOX / siPD-L1的抗肿瘤作用。NPDOX / siPD-L1在肝癌中诱导的免疫原性细胞死亡（ICD）和PD-L1过表达的治疗。体内研究表明，在H22荷瘤动物模型中，静脉注射NPDOX / siPD-L1可显着抑制肿瘤体积和肿瘤组织的PD-L1表达。此外，NPDOX / siPD-L1的治疗还调节了肿瘤组织中成熟的树突状细胞和细胞毒性T细胞的数量以及细胞因子的产生。两者合计，NPDOX / siPD-L1通过诱导ICD和抑制PD-L1过表达，对肝癌表现出显着的抗肿瘤作用。NPDOX / siPD-L1的治疗还调节了肿瘤组织中成熟树突状细胞和细胞毒性T细胞的数量以及细胞因子的产生。两者合计，NPDOX / siPD-L1通过诱导ICD和抑制PD-L1过表达，对肝癌表现出显着的抗肿瘤作用。NPDOX / siPD-L1的治疗还调节了肿瘤组织中成熟树突状细胞和细胞毒性T细胞的数量以及细胞因子的产生。两者合计，NPDOX / siPD-L1通过诱导ICD和抑制PD-L1过表达，对肝癌表现出显着的抗肿瘤作用。