葛根来源外泌体囊泡在结肠癌类器官及细胞模型中的作用  

Role of P.lobata root-derived exosome-like nanovesicles in colorectal cancer organoids and cell models

作　　者：闫芳[1] 黄世远 王嘉 黄伟[4] 邱怡 刘涛[2] 杨洋 杨健 YAN Fang;HUANG Shiyuan;WANG Jia;HUANG Wei;QIU Yi;LIU Tao;YANG Yang;YANG Jian(Geriatric Diseases Institute of Chengdu,Department of Geriatrics,Center for Medicine Research and Translation,Chengdu Fifth People's Hospital,Sichuan Chengdu 611130,China;Sichuan Industrial Institute of Antibiotics,School of Pharmacy,Chengdu University,Sichuan Chengdu 610106,China;Chengdu Organoidmed Medical Laboratory Co.,Ltd.,Sichuan Chengdu 610041,China;Traditional Chinese Medicine Department,Chengdu Fifth People's Hospital,Sichuan Chengdu 611130,China;Department of Gastroenterology,Sir Run Run Hospital,Nanjing Medical University,Jiangsu Nanjing 211100,China;General Surgery Department,Chengdu Fifth People's Hospital,Sichuan Chengdu 611130,China)

机构地区：[1]成都市老年疾病研究所,成都市第五人民医院老年医学科,成都市第五人民医院医学研究与转化中心四川成都611130 [2]成都大学药学院,四川抗菌素研究所,四川成都610106 [3]成都诺医德医学检验实验室有限公司,四川成都610041 [4]成都市第五人民医院中医科,四川成都611130 [5]南京医科大学附属逸夫医院消化内科,江苏南京211100 [6]成都市第五人民医院普通外科,四川成都611130

出　　处：《现代肿瘤医学》2025年第6期902-910,共9页Journal of Modern Oncology

基　　金：四川省科技厅项目(编号:2022YFS0165);成都中医药大学院校联合创新项目(编号:LH202402002);成都医学研究项目(编号:2023459,2024107);成都中医药大学杏林学者(编号:XJ2023001102)。

摘　　要：目的:在结肠癌细胞系和结肠癌类器官上探讨新鲜植物葛根来源的外泌体样囊泡(fresh P.lobata root-derived exosome-like nanovesicles,P-ELNs)的抗肿瘤效果。方法:HT-29、Caco-2结肠癌细胞系和HUVEC非肿瘤细胞系中加入不同浓度的P-ELNs,使用CellTiter-Lumi^(TM)发光法细胞活力检测试剂盒检测细胞活力,AO/PI细胞双染进行细胞活死染色,Caspase3/7荧光探针来检测细胞凋亡情况;构建结肠癌患者组织来源的结肠癌类器官,H&E染色观察类器官特征,不同浓度的P-ELNs处理后AO/PI细胞双染进行细胞活死染色,类器官活性检测试剂盒检测类器官活力。结果:细胞活力检测结果显示,P-ELNs以浓度依赖的方式选择性抑制癌细胞增殖,390μg/mL的P-ELNs处理后,Caco-2的敏感性较低,细胞存活率为47.8%,更为敏感的HT-29的细胞存活率仅为25.82%,而HUVEC的细胞存活率为64.8%;细胞活死染色显示,P-ELNs处理后活细胞的数量相较于完全培养基处理的对照显著降低,同时P-ELNs处理后死细胞数量显著增多,Caspase3/7荧光探针来检测凋亡发现P-ELNs处理后未发现明显的细胞凋亡;H&E染色观察肿瘤类器官形态,发现其具有与其来源肿瘤组织相似的特征,AO/PI细胞活死染色结果显示,P-ELNs处理后存活的类器官数量相较于对照组显著降低,且出现了大量的类器官死亡,类器官的增殖实验结果显示在用梯度浓度的P-ELNs处理类器官后,类器官表现出浓度依赖性的活性降低,且相较于细胞系敏感性更高。结论:P-ELNs能够通过非凋亡途径诱导结肠癌细胞死亡来抑制其增殖,同样地P-ELNs能够诱导细胞的死亡来抑制结肠癌类器官增殖,并呈现浓度依赖性。Objective:To investigate the antitumor effects of fresh P.lobata root-derived exosome-like nanovesicles(P-ELNs)in colon cancer cell lines and organoids.Methods:HT-29,Caco-2 colon cancer cell line and HUVEC non-tumor cell lines were added with different concentrations of P-ELNs.Cell viability was detected using CellTiter-Lumi^(TM)luminescence cell viability assay kit,and cell viability was stained by AO/PI double staining.Caspase3/7 fluorescent probe was used to detect cell apoptosis.Constructing organoids derived from tissues of patients with colon cancer,and organoid characteristics were observed by H&E staining.After different concentrations of P-ELNs were treated,AO/PI cells were double-dyed for cell survival and death staining.Organoid activity detection kit was used to detect organoid activity.Results:The results of the cell viability assay demonstrated that P-ELNs selectively inhibited cancer cell proliferation in a concentration-dependent manner.Following treatment with P-ELNs at a concentration of 390μg/mL,the survival rate of Caco-2 cells decreased with 47.8%,and the survival rate of HT-29 cells was reduced to 25.82%.In contrast,the survival rate of HUVEC cells was 64.8%.Staining for cell death indicated a significant reduction in the number of viable cells and a significant increase in the number of dead cells in the P-ELNs treated group compared to the control group,which was treated with complete medium.Additionally,caspase-3/7 fluorescence was observed.Caspase3/7 fluorescent probe was employed to assess cell apoptosis,revealing no significant apoptosis following P-ELNs treatment.Hematoxylin and eosin(HE)staining was utilized to examine the morphology of tumor organoids,which exhibited characteristics similar to the original tumor tissues.Acridine orange/propidium iodide(AO/PI)staining indicated a significantly reduced number of surviving organoids post P-ELNs treatment compared to the control group,with a substantial number of organoids undergoing cell death.The proliferation assay demonstrated that

关 键 词：葛根来源的外泌体样囊泡 结肠癌类器官 结肠癌 药物筛选 

分 类 号：R735.3[医药卫生—肿瘤]