细胞外囊泡装载多柔比星对人膀胱癌BIU-87细胞增殖及凋亡的影响  

Effect of Extracellular Vesicles Loaded with Doxorubicin on the Proliferation and Apoptosis of Human Bladder Cancer BIU-87 Cells

作　　者：马燕凌 晏菲 魏武杰 邓洁 李黎 刘莉 孙建海 MA Yanling;YAN Fei;WEI Wujie;DENG Jie;LI Li;LIU Li;SUN Jianhai(Department of Oncology,Hubei No.3 People's Hospital of Jianghan University,Wuhan,Hubei,China 430000)

机构地区：[1]江汉大学附属湖北省第三人民医院肿瘤科,湖北武汉430000

出　　处：《中国药业》2022年第14期60-64,共5页China Pharmaceuticals

基　　金：湖北省卫生健康委科研立项项目[WJ2019F184]。

摘　　要：目的探讨细胞外囊泡(EVs)装载多柔比星对人膀胱癌BIU-87细胞增殖及凋亡的影响。方法体外培养BIU-87细胞并进行EVs提取,制备装载多柔比星的载药囊泡。实验分为空白对照组(不进行处理)、EVs组(EVs混悬液,100个/细胞)、多柔比星组(0.3μg多柔比星)、载药囊泡组(载药囊泡混悬液,100个/细胞)。采用MTT法和流式细胞法分别检测细胞增殖率和凋亡率,实时荧光聚合酶链式反应(RT-PCR)法和Western blot法分别检测细胞中糖原合成酶激酶-3β(GSK-3β)、β-连环蛋白(β-catenin)、Notch1的mRNA及蛋白表达水平。结果EVs组和载药囊泡组细胞内PKH67标记的团块状绿色荧光均匀;与空白对照组比较,EVs组上述指标均无显著差异(P>0.05);与空白对照组及EVs组比较,多柔比星组和载药囊泡组细胞增殖率、β-catenin和Notch1 mRNA及蛋白表达水平均显著降低,细胞凋亡率、GSK-3βmRNA及蛋白表达水平均显著升高,且载药囊泡组更优(P<0.05)。结论EVs装载多柔比星抑制BIU-87细胞增殖和促进其凋亡的作用较单用多柔比星强,机制可能与抑制β-catenin/Notch1信号通路有关。Objective To investigate the effect of extracellular vesicles(EVs)loaded with doxorubicin on the proliferation and apoptosis of human bladder cancer BIU-87 cells.Methods BIU-87 cells were cultured in vitro and EVs were extracted to prepare drug-loaded vesicles loaded with doxorubicin.The blank control group(without treatment),EVs group(EVs suspension,100 EVs/cell),doxorubicin group(0.3μg of doxorubicin)and drug-loaded vesicle group(drug-loaded vesicle suspension,100 EVs/cells)were set up in this experiment.MTT assay and flow cytometry were used to detect the proliferation rate and apoptosis rate of cells,and real-time fluorescence polymerase chain reaction(RT-PCR)and Western blot were used to detect the expression levels of glycogen synthase kinase-3β(GSK-3β),β-catenin and Notch1 mRNA and protein in cells.Results In the EVs group and drug-loaded vesicle group,the PKH67 labeled green fluorescence in BIU-87 cells was uniform.Compared with those in the blank control group,there was no significant difference in the above indexes in the EVs group(P>0.05).Compared with those in the blank control group and EVs group,the cell proliferation rate,the expression levels ofβ-catenin and Notch1 mRNA and protein were significantly lower,while the apoptosis rate and the expression levels of GSK-3βmRNA and protein were significantly higher in the doxorubicin group and drug-loaded vesicle group,and those in the drug-loaded vesicle group were better(P<0.05).Conclusion EVs loaded with doxorubicin has a stronger effect on inhibiting the proliferation of BIU-87 cells and promoting the apoptosis of BIU-87 cells than that of doxorubicin,and its mechanism may be related to the inhibition of the expression ofβ-catenin/Notch1 signaling pathway.

关 键 词：细胞外囊泡 多柔比星 膀胱癌 细胞增殖 细胞凋亡 β-catenin/Notch1通路 

分 类 号：R979.14[医药卫生—药品] R96[医药卫生—药学]