目的:探明RAC1蛋白調(diào)控阿霉素（DOX）誘導(dǎo)腫瘤轉(zhuǎn)移機(jī)制,設(shè)計(jì)靶向RAC1的小干擾RNA（siRNA）遞釋系統(tǒng),抑制DOX治療誘導(dǎo)的腫瘤皮質(zhì)間質(zhì)樣（EMT）轉(zhuǎn)化。創(chuàng)新點(diǎn):探明了DOX通過RAC1蛋白誘導(dǎo)腫瘤細(xì)胞發(fā)生轉(zhuǎn)移性變化的機(jī)制,構(gòu)建了靶向沉默RAC1蛋白的siRNA遞釋系統(tǒng),有效降低了DOX化療后腫瘤組織轉(zhuǎn)移的風(fēng)險(xiǎn)。方法:選用MCF-7細(xì)胞為模型細(xì)胞,體外考察DOX相關(guān)活性氧（ROS）對(duì)RAC1蛋白的調(diào)控作用。針對(duì)腫瘤細(xì)胞內(nèi)高谷胱甘肽（GSH）濃度的氧化還原條件,選用二硫鍵為敏感橋鏈,殼聚糖為親水性骨架,硬脂胺為疏水基團(tuán),構(gòu)建氧化還原敏感型基因藥物載體CSO-ss-SA。選擇靶向沉默RAC1的siRNA為模型藥物,構(gòu)建基因藥物遞釋系統(tǒng)CSO-ss-SA/siRNA�？疾霤SO-ss-SA/siRNA的理化性質(zhì)、體外敏感釋放及細(xì)胞攝取。通過免疫熒光染色法、蛋白免疫印跡法和細(xì)胞侵襲性實(shí)驗(yàn)考察基因藥物遞釋系統(tǒng)的細(xì)胞藥效。選用Balb/c裸鼠構(gòu)建MCF-7乳腺腫瘤原位荷瘤動(dòng)物模型,考察CSO-ss-SA/siRNA對(duì)DOX治療誘導(dǎo)腫瘤組織EMT轉(zhuǎn)化的抑制作用。結(jié)論:RAC1是DOX誘導(dǎo)細(xì)... 

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

【文章頁數(shù)】：16 頁

【文章目錄】：
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


【參考文獻(xiàn)】：
期刊論文
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[2]多柔比星脂質(zhì)體在乳腺癌輔助化療中的臨床應(yīng)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)



本文編號(hào)：3081941