基于外泌体-串联siRNA自组装靶系统在KRAS驱动型胰腺癌治疗中的应用  

The application of exosome-based tandem siRNA self-assembled targeting system in the treatment of KRAS-driven pancreatic cancer

作　　者：许烨 詹守斌 周圣凯 陈熹[1] XU Ye;ZHAN Shou-bin;ZHOU Sheng-kai(Nanjing University,Nanjing 210033,China)

机构地区：[1]南京大学,210033

出　　处：《中国实用医药》2025年第4期171-176,共6页China Practical Medicine

摘　　要：目的探讨外泌体-串联小分子干扰核糖核酸(siRNA)自组装靶系统在鼠类肉瘤病毒癌基因(KRAS)驱动型胰腺癌治疗中的应用效果。方法整合外泌体靶向元件PTP-Lamp2B及串联siRNA表达元件,成功构建了一种新型siRNA自组装系统。首先在胰腺癌细胞系中筛选有效的siRNA并进行串联,接着对细胞培液产生提取的外泌体的KRAS基因沉默的有效性、PTP的靶向性、对胰腺癌细胞增殖的抑制程度进行验证。主要通过检测不同处理组KRAS蛋白的表达情况以评判效果。结果相比于对照组,CMV-siR-662和CMV-siR-601基因环路处理后KRAS的表达下降1.5~3倍多(P<0.0001);串联后表达的基因环路处理后,PANC-1细胞系中KRAS表达下降18倍(P<0.0001),MIA PaCa-2细胞系中KRAS表达下降140多倍(P<0.0001);串联后的基因环路使胰腺癌细胞增殖速度减缓1倍(P<0.0001);在基因环路中额外加装靶向肽后产生的外泌体,到达胰腺癌细胞中的能力显著上升,是无靶向肽外泌体的5倍左右(P<0.0001)。最后,整合靶向元件PTP-Lamp 2B及串联siRNA表达元件的基因环路CMV-PTP-siRmix,产生的靶向性外泌体可以显著抑制胰腺癌细胞中KRAS的表达、下游信号通路中的磷酸化的细胞外调节蛋白激酶(ERK)的表达、胰腺癌细胞的增殖。结论该外泌体-串联siRNA自组装靶系统在KRAS驱动型胰腺癌治疗中具有潜在的应用前景。Objective To explore the application effect of exosome-based tandem small interfering ribonucleic acid(siRNA)self-assembled targeting system in the treatment of kirsten rat sarcoma viral oncogene(KRAS)-driven pancreatic cancer.Methods The exosome targeting element PTP-Lamp 2B and tandem siRNA expression elements were integrated to construct a novel siRNA self-assembling system.Initially,effective siRNAs were screened in pancreatic cancer cell lines and then tandemly linked.Subsequently,the efficacy of exosomes extracted from cell culture supernatants in silencing KRAS gene,PTP targeting,and the extent of inhibition of pancreatic cancer cell proliferation were verified.The main evaluation was based on the expression of KRAS protein in different treatment groups.Results Compared to the control group,the expression of KRAS decreased by 1.5-3 times after treatment with the CMV-siR-662 and CMV-siR-601 gene circuits(P<0.0001).The tandem expression of these gene circuits further enhanced this inhibitory effect,with KRAS expression in the PANC-1 cell line decreasing by 18 times(P<0.0001),and in the MIA PaCa-2 cell line by more than 140 times(P<0.0001).The tandem gene circuit slowed down the proliferation rate of pancreatic cancer cells by 1 time(P<0.0001).The exosomes produced after the addition of a targeting peptide to the gene circuit significantly increased their ability to reach pancreatic cancer cells,which was about 5 times that of exosomes without a targeting peptide(P<0.0001).Finally,the gene circuit CMV-PTP-siRmix,integrating the targeting element PTP-Lamp 2B and tandem siRNA expression elements,produced targeted exosomes that significantly inhibited the expression of KRAS in pancreatic cancer cells,the expression of phosphorylated extracellular regulated protein kinases(ERK)in the downstream signaling pathway,and the proliferation of pancreatic cancer cells.Conclusion This exosome-based tandem siRNA self-assembled targeting system has potential application prospects in the treatment of KRAS-driven pancreatic c

关 键 词：驱动型胰腺癌 鼠类肉瘤病毒癌基因 外泌体-串联小分子干扰核糖核酸 自组装靶系统 靶向治疗 

分 类 号：R73[医药卫生—肿瘤]