RAC1蛋白靶向氧化还原敏感型siRNA递释系统抑制化疗诱导乳腺肿瘤转移的研究（英文）

发布时间：2021-03-14 09:46

　　目的:探明RAC1蛋白调控阿霉素（DOX）诱导肿瘤转移机制,设计靶向RAC1的小干扰RNA（siRNA）递释系统,抑制DOX治疗诱导的肿瘤皮质间质样（EMT）转化。创新点:探明了DOX通过RAC1蛋白诱导肿瘤细胞发生转移性变化的机制,构建了靶向沉默RAC1蛋白的siRNA递释系统,有效降低了DOX化疗后肿瘤组织转移的风险。方法:选用MCF-7细胞为模型细胞,体外考察DOX相关活性氧（ROS）对RAC1蛋白的调控作用。针对肿瘤细胞内高谷胱甘肽（GSH）浓度的氧化还原条件,选用二硫键为敏感桥链,壳聚糖为亲水性骨架,硬脂胺为疏水基团,构建氧化还原敏感型基因药物载体CSO-ss-SA。选择靶向沉默RAC1的siRNA为模型药物,构建基因药物递释系统CSO-ss-SA/siRNA。考察CSO-ss-SA/siRNA的理化性质、体外敏感释放及细胞摄取。通过免疫荧光染色法、蛋白免疫印迹法和细胞侵袭性实验考察基因药物递释系统的细胞药效。选用Balb/c裸鼠构建MCF-7乳腺肿瘤原位荷瘤动物模型,考察CSO-ss-SA/siRNA对DOX治疗诱导肿瘤组织EMT转化的抑制作用。结论:RAC1是DOX诱导细... 

【文章来源】：Journal of Zhejiang University-Science B（Biomedicine & Biotechnology）. 2020,21(03)

【文章页数】：16 页

【文章目录】：
1 Introduction
2 Materials and methods
    2.1 Materials
    2.2 Cell culture
    2.3 Cell invasion assay
    2.4 ROS determination
    2.5 Immunofluorescence staining of RAC1 and cytoskeleton
    2.6 Western blot assay for RAC1 expression
    2.7 Synthesis and characterization of CSO-ss-SA
    2.8 Preparation and characterization of CSO-ss-SA/siRNA
    2.9 Gel electrophoresis experiments
    2.1 0 Cellular uptake
    2.1 1 Inhibition of RAC1 protein and cytoskeleton by CSO-ss-SA/siRNA
    2.1 2 Cytotoxicity of CSO-ss-SA/siRNA
    2.1 3 Inhibition of DOX-induced cell invasion by CSO-ss-SA/siRNA in vitro
    2.1 4 Inhibition of DOX-induced tumor EMT by CSO-ss-SA/siRNA in vivo
    2.1 5 Statistical analysis
3 Results
    3.1 Chemotherapy-induced tumor cell invasive-ness and ROS increase
    3.2 Chemotherapy-induced RAC1 expression and cytoskeleton reconstruction
    3.3 Synthesis and structure verification of CSO-ss-SA
    3.4 Physicochemical characteristics of CSO-ss-SA
    3.5 Preparation and characterization of CSO-ss-SA/siRNA complexes
    3.6 GSH-triggered siRNA release
    3.7 Cellular uptake of CSO-ss-SA/siRNA
    3.8 Inhibition of RAC1 by CSO-ss-SA/siRNA in vitro
    3.9 Cytotoxicity of CSO-ss-SA/siRNA complexes combined with DOX in vitro
    3.1 0 Therapy efficiency of CSO-ss-SA/siRNA com-plexes combined with DOX in vivo
4 Discussion
5 Conclusions
Contributors
Compliance with ethics guidelines


【参考文献】：
期刊论文
[1]脂筏标记蛋白flotillins在肿瘤中的研究进展（英文）[J]. Xu-xu LIU,Wei-dong LIU,Lei WANG,Bin ZHU,Xiao SHI,Zi-xuan PENG,He-cheng ZHU,Xing-dong LIU,Mei-zuo ZHONG,Dan XIE,Mu-sheng ZENG,Cai-ping REN.  Journal of Zhejiang University-Science B（Biomedicine & Biotechnology）. 2018(03)
[2]多柔比星脂质体在乳腺癌辅助化疗中的临床应用（英文）[J]. Ming ZHAO,Xian-feng DING,Jian-yu SHEN,Xi-ping ZHANG,Xiao-wen DING,Bin XU.  Journal of Zhejiang University-Science B（Biomedicine & Biotechnology）. 2017(01)



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