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壳聚糖包被的阿霉素纳米颗粒药物递送系统通过p53/PRC1途径抑制肝癌细胞生长。

Chitosan-coated doxorubicin nano-particles drug delivery system inhibits cell growth of liver cancer via p53/PRC1 pathway.

作者信息

Ye Bai-Liang, Zheng Ru, Ruan Xiao-Jiao, Zheng Zhi-Hai, Cai Hua-Jie

机构信息

Department of General Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, Zhejiang, China.

Department of General Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, Zhejiang, China.

出版信息

Biochem Biophys Res Commun. 2018 Jan 1;495(1):414-420. doi: 10.1016/j.bbrc.2017.10.156. Epub 2017 Oct 31.

Abstract

BACKGROUND

Nano-particles have been widely used in target-specific drug delivery system and showed advantages in cancers treatment. This study aims to evaluate the effect of chitosan coated doxorubicin nano-particles drug delivery system in liver cancer.

METHODS

The chitosan nano-particles were prepared by using the ionic gelation method. The characterizations of the nano-particles were determined by transmission electron microscopy. The cytotoxicity was detected by MTT assay, and the endocytosis, cell apoptosis and cell cycle were examined by flow cytometry. The protein level was analyzed with western blot. The dual luciferase reporter assay was performed to assess the interaction between p53 and the promoter of PRC1, and chromatin immune-precipitation was used to verify the binding between them.

RESULTS

The FA-CS-DOX nano-particles were irregular and spherical particles around 30-40 nm, with uniform size and no adhesion. No significant difference was noted in doxorubicin release rate between CS-DOX and FA-CS-DOX. FA-CS-DOX nano-particles showed stronger cytotoxicity than CS-DOX. FA-CS-DOX nano-particles promoted the apoptosis and arrested cell cycle at G2/M phase, and they up-regulated p53. FA-CS-DOX nano-particles inhibited cell survival through p53/PRC1 pathway.

CONCLUSION

Chitosan-coated doxorubicin nano-particles drug delivery system inhibits cell growth of liver cancer by promoting apoptosis and arresting cell cycle at G2/M phase through p53/PRC1 pathway.

摘要

背景

纳米粒子已广泛应用于靶向给药系统,并在癌症治疗中显示出优势。本研究旨在评估壳聚糖包被阿霉素纳米粒子给药系统对肝癌的影响。

方法

采用离子凝胶法制备壳聚糖纳米粒子。通过透射电子显微镜对纳米粒子进行表征。采用MTT法检测细胞毒性,通过流式细胞术检测内吞作用、细胞凋亡和细胞周期。用蛋白质免疫印迹法分析蛋白质水平。进行双荧光素酶报告基因检测以评估p53与PRC1启动子之间的相互作用,并用染色质免疫沉淀法验证它们之间的结合。

结果

FA-CS-DOX纳米粒子为不规则球形颗粒,粒径约30-40nm,大小均匀,无粘连。CS-DOX和FA-CS-DOX之间的阿霉素释放率无显著差异。FA-CS-DOX纳米粒子比CS-DOX表现出更强的细胞毒性。FA-CS-DOX纳米粒子促进细胞凋亡并使细胞周期停滞在G2/M期,且上调p53。FA-CS-DOX纳米粒子通过p53/PRC1途径抑制细胞存活。

结论

壳聚糖包被阿霉素纳米粒子给药系统通过p53/PRC1途径促进细胞凋亡并使细胞周期停滞在G2/M期,从而抑制肝癌细胞生长。

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