松果菊苷下调Skp2的表达抑制胶质母细胞瘤上皮间质转化及胶质瘤干细胞干性  

Echinacoside inhibits Skp2-induced epithelial-mesenchymal transition and stemness in glioblastoma

作　　者：吴振德 王兆涛 鞠其超 张波 WU Zhen-de;WANG Zhao-tao;JU Qi-chao;ZHANG Bo(Department of Neurosurgery,Xiamen Chang Gung Hospital,Xiamen Fujian 361028;Department of Neurosurgery,the Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 560210)

机构地区：[1]厦门长庚医院神经外科,福建厦门361028 [2]广州医科大学附属第二医院神经外科,广州560210

出　　处：《中南药学》2024年第3期626-632,共7页Central South Pharmacy

基　　金：广东省自然科学基金面上项目(No.2019A1515010926)。

摘　　要：目的研究松果菊苷(ECH)对胶质母细胞瘤(GBM)的上皮间质转化(EMT)及其对GBM干细胞的影响,并探讨其分子机制。方法①体外实验:将GBM U87和U251细胞以0、5、10和20μmol·L^(-1) ECH浓度处理24 h。采用CCK-8法评估细胞活性;进行克隆形成实验以测定克隆能力;通过Transwell实验评估迁移和侵袭能力;利用球形成实验评价GBM干细胞的成球能力;通过细胞免疫荧光检测干细胞标志物;使用Western blot分析EMT和干细胞相关蛋白表达。②体内实验:在裸鼠皮下注射U87细胞,每组5只。实验组腹腔注射ECH,对照组注射等体积溶剂。最后测定肿瘤体积、重量和体重,并用Western blot分析肿瘤中的Skp2、EMT和干细胞相关蛋白表达。结果①体外实验显示,相较于0μmol·L^(-1) ECH组,10、20μmol·L^(-1) ECH处理显著降低了细胞活性(P<0.05);在5、10和20μmol·L^(-1) ECH组中克隆形成数量显著减少(P<0.05);迁移和侵袭能力在5、10μmol·L^(-1) ECH组中随浓度升高而降低(P<0.05);成球能力和干细胞标志物表达及Skp2、EMT和干细胞特性相关蛋白表达在5、10μmol·L^(-1) ECH组中均显著降低(P<0.05)。②体内实验显示,ECH处理小鼠的肿瘤体积和重量显著小于对照组(P<0.05),且Skp2、EMT(Vimentin、Snail)和干细胞标志物(Sox2、Nestin)的表达显著减少(P<0.05);两组小鼠体重差异无统计学意义(P>0.05)。结论ECH通过降低Skp2表达抑制GBM的EMT和干细胞特性。Objective To determine the impact of echinacoside(ECH)on the epithelial-mesenchymal transition(EMT)and glioma stem cell characteristics in glioblastoma,and to elucidate related molecular mechanism.Methods①In vitro experiments:glioblastoma U87 and U251 cells were cultured and treated with 0,5,10,and 20μmol·L^(-1) ECH for 24 hours;the cell viability was evaluated with the CCK-8 assay;clonogenic capacity was determined by the colony formation assay,migration and invasion by transwell assay,and stem cell potential by sphere formation assay.The expression of stem cell markers was assessed with immunofluorescence;EMT and stemnessrelated protein expressions were analyzed with Western blot.②In vivo experiments:U87 cells were subcutaneously implanted into nude mice(five mice in each group).The experimental group was administered daily intraperitoneal injections of ECH,while the control group received a corresponding volume of the vehicle.At the end of the experiment,tumor volume,mouse weight,tumor weight were measured,and Skp2,EMT,and stemness-related protein expressions in tumors were analyzed by Western blot.Results①In vitro experiments showed that,compared to the 0μmol·L^(-1) ECH group,the 10 and 20μmol·L^(-1) ECH groups significantly reduced cell viability(P<0.05);the number of colonies formed was significantly decreased in the 5,10,and 20μmol·L^(-1) ECH groups(P<0.05);migration and invasion abilities were reduced in the 5 and 10μmol·L^(-1) ECH groups(P<0.05);spheroid formation ability and stem cell marker expression,Skp2 and EMT expression,and stem cell characteristic proteins significantly decreased in the 5 and 10μmol·L^(-1) ECH groups(P<0.05).②In vivo experiments showed that,tumor volume and weight in mice treated with ECH were obviously smaller than those in the control group(P<0.05),and the expression of Skp2 and EMT markers(Vimentin,Snail),and stem cell markers(Sox2,Nestin)significantly decreased;there was no significant difference in the body weight between both groups.Conclusion ECH inhibi

关 键 词：胶质母细胞瘤 松果菊苷 上皮间质转化 干细胞干性 SKP2 

分 类 号：R285.5[医药卫生—中药学]