脂质体共递送槲皮素与多柔比星通过调控上皮间质转化过程抑制视网膜母细胞瘤的研究

Liposome co-delivery of quercetin and doxorubicin in inhibiting retinoblastoma by regulating epithelial-mesenchymal transition process

调控上皮-间质转化(epithelial-mesenchymal transition,EMT)过程是抑制肿瘤生长和转移的重要策略。该研究基于视网膜母细胞瘤的EMT过程,构建同时负载槲皮素(quercetin,QUE)与多柔比星(doxorubicin,DOX)的脂质体,用于研究QUE与DOX联用对视网膜母细胞瘤的治疗作用及其机制。通过单因素考察对槲皮素与多柔比星双载脂质体(QD Lipo)进行处方工艺优化,最终得到球形,粒径(108.87±1.93)nm,PDI 0.13±0.02,Zeta电位(-34.83±1.92)mV的双载脂质体,其中QUE、DOX包封率分别为96.20%±4.40%、91.17%±4.41%。以人视网膜母细胞瘤Y79作为体外细胞模型,采用共聚焦显微镜证明QD Lipo能提高Y79对其摄取效率。采用CCK-8法证明QUE∶DOX在1∶1~1∶2时具有最佳联合治疗作用。流式细胞技术证明QD Lipo增强了诱导Y79细胞凋亡的能力。通过Western blot技术发现QD Lipo能显著降低EMT通路相关蛋白vimentin和α-SMA的表达,并且荧光法检测到治疗后的Y79细胞内ROS水平显著降低。以上结果显示,脂质体共递送QUE与DOX可提高药物向视网膜母细胞瘤的输送效率,通过下调ROS水平抑制视网膜母细胞瘤的EMT过程,同时增强DOX对视网膜母细胞瘤的杀伤作用。

Regulating the process of epithelial-mesenchymal transition(EMT)is an essential strategy to inhibit tumor growth and metastasis.This study is based on the EMT process of retinoblastoma and constructs quercetin(QUE)and doxorubicin(DOX)co-loaded liposome(QD Lipo)to investigate the therapeutic effect and mechanisms of combined QUE and DOX treatment on retinoblastoma.Single-factor experiments were conducted to optimize the prescription process of QD Lipo.Eventually,spherical particles with a diameter of(108.87±1.93)nm,a PDI of 0.13±0.02,and a Zeta potential of(-34.83±1.92)mV were obtained.The encapsulation rates of QUE and DOX were 96.20%±4.40%and 91.17%±4.41%,respectively.Y79 human retinoblastoma cells were used as an in vitro cellular model,and confocal microscopy demonstrated that QD Lipo could enhance Y79 uptake efficiency.The CCK-8 assay confirmed that the optimal combination therapy effect of QUE and DOX occurred at a mass ratio of 1∶1 to 1∶2.Flow cytometry showed that QD Lipo enhanced the induction of apoptosis in Y79 cells.Western blot analysis revealed that QD Lipo significantly reduced the expression of EMT pathway-related proteins vimentin andα-SMA.Fluorescence assays detected a significant decrease in ROS levels in Y79 cells after treatment with QD.These results indicated that liposomal co-delivery of QUE and DOX can enhance drug delivery efficiency to retinoblastoma cells,inhibit the EMT process in retinoblastoma by downregulating ROS levels,and enhance the cytotoxicity of DOX against retinoblastoma.