Abstract
Metastasis is one of the main reasons causing death in cancer patients. It was reported that chemotherapy might induce metastasis. In order to uncover the mechanism of chemotherapy-induced metastasis and find solutions to inhibit treatment-induced metastasis, the relationship between epithelial-mesenchymal transition (EMT) and doxorubicin (DOX) treatment was investigated and a redox-sensitive small interfering RNA (siRNA) delivery system was designed. DOX-related reactive oxygen species (ROS) were found to be responsible for the invasiveness of tumor cells in vitro, causing enhanced EMT and cytoskeleton reconstruction regulated by Ras-related C3 botulinum toxin substrate 1 (RAC1). In order to decrease RAC1, a redox-sensitive glycolipid drug delivery system (chitosan-ss-stearylamine conjugate (CSO-ss-SA)) was designed to carry siRNA, forming a gene delivery system (CSO-ss-SA/siRNA) down-regulating RAC1. CSO-ss-SA/siRNA exhibited an enhanced redox sensitivity compared to nonresponsive complexes in 10 mmol/L glutathione (GSH) and showed a significant safety. CSO-ss-SA/siRNA could effectively transmit siRNA into tumor cells, reducing the expression of RAC1 protein by 38.2% and decreasing the number of tumor-induced invasion cells by 42.5%. When combined with DOX, CSO-ss-SA/siRNA remarkably inhibited the chemotherapy-induced EMT in vivo and enhanced therapeutic efficiency. The present study indicates that RAC1 protein is a key regulator of chemotherapy-induced EMT and CSO-ss-SA/siRNA silencing RAC1 could efficiently decrease the tumor metastasis risk after chemotherapy.
摘要
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 诱导细胞侵袭性的关键蛋白. 靶向沉默 RAC1 的siRNA 基因递释系统可在体内外有效抑制 DOX 治疗诱导的肿瘤组织EMT 转化, 降低化疗诱导转移的风险.
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Change history
28 February 2022
An Erratum to this paper has been published: https://doi.org/10.1631/jzus.B19e0468
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Xuan LIU performed the experimental research and data analysis, wrote and edited the manuscript. Xue-qing ZHOU performed the data analysis and participated in the paperwork. Xu-wei SHANG contributed in polymer synthesis. Li WANG contributed in image analysis. Yi LI collected and analyzed the data. Hong YUAN and Fu-qiang HU contributed in the study design. All authors have read and approved the final manuscript, have full access to all the data in the study, and take responsibility for the integrity and security of the data.
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Xuan LIU, Xue-qing ZHOU, Xu-wei SHANG, Li WANG, Yi LI, Hong YUAN, and Fu-qiang HU declare that they have no conflict of interest.
All institutional and national guidelines for the care and use of laboratory animals were followed.
Project supported by the National Natural Science Foundation of China (No. 81773648), the Zhejiang Provincial Natural Science Foundation of China (No. D19H30001), and the Chinese Postdoc Funding (No. 2018M630686)
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Liu, X., Zhou, Xq., Shang, Xw. et al. Inhibition of chemotherapy-related breast tumor EMT by application of redox-sensitive siRNA delivery system CSO-ss-SA/siRNA along with doxorubicin treatment. J. Zhejiang Univ. Sci. B 21, 218–233 (2020). https://doi.org/10.1631/jzus.B1900468
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DOI: https://doi.org/10.1631/jzus.B1900468
Key words
- Doxorubicin
- Tumor metastasis
- Ras-related C3 botulinum toxin substrate 1 (RAC1)
- Epithelial-mesenchymal transition (EMT)
- Chitosan micelles
- Small interfering RNA (siRNA)