槲皮素对肺癌干细胞活性的影响  

Effects of quercetin on the activity of lung cancer stem cells

作　　者：朱剑云 谢春凤[2] 钟才云[2] Zhu Jianyun;Xie Chunfeng;Zhong Caiyun(Department of Laboratory,Suzhou Municipal hospital,The Affiliated Suzhou Hospital of Nanjing Medical University,Suzhou 215008,Jiangsu China;Department of Nutrition and Food Safety,School of Public Health,Nanjing Medical University,Nanjing 211166,Jiangsu China)

机构地区：[1]南京医科大学附属苏州医院苏州市立医院北区中心实验室,江苏苏州215008 [2]南京医科大学公共卫生学院营养与食品卫生学系,南京211166

出　　处：《肿瘤代谢与营养电子杂志》2022年第5期616-623,共8页Electronic Journal of Metabolism and Nutrition of Cancer

基　　金：江苏省科协青年科技人才托举工程项目(TJ-2021-029);苏州市“姑苏卫生人才计划”青年拔尖人才项目(GSWS2020075)。

摘　　要：目的 探索槲皮素对A549肺癌干细胞活性及对Wnt/β-连环蛋白(β-catenin)和音猬因子(sonic hedgehog,SHH)信号通路的影响。方法 使用不同浓度槲皮素(0、5、10、25、50、100和200μmol/L)处理肺癌A549细胞24、48和72 h,MTT法检测细胞存活率。采用无血清培养法分离富集肺癌干细胞,观察槲皮素对A549悬浮细胞球体积、数量以及肺癌干细胞分子标志物表达的影响;流式细胞术检测细胞凋亡;蛋白质印迹法检测增殖凋亡相关指标以及Wnt/β-catenin和SHH信号通路关键分子的表达变化。结果 槲皮素能够显著抑制A549细胞活力,且呈现时间和浓度依赖性;与空白对照组相比,槲皮素均显著抑制A549成球能力和肺癌干细胞分子标志物[CD133、CD44和醛脱氢酶1A1(ALDH1A1)]表达;同时,肺癌干细胞增殖相关蛋白增殖细胞核抗原(PCNA)、细胞周期蛋白D1(Cyclin D1)和B细胞白血病/淋巴瘤-2(Bcl2)水平显著下调,凋亡相关指标Bcl2相关蛋白X(Bax)和半胱氨酰天冬氨酸特异性蛋白酶-3(Cleaved Caspase 3)表达显著上调;槲皮素处理后,肺癌干细胞中Wnt/β-catenin和SHH信号通路关键分子磷酸化糖原合成酶激酶-3β(p-GSK3β)(Ser9)、β-catenin、原癌基因蛋白c-Myc、胶质瘤相关癌基因同源物1(GLi1)、GLi2和SHH的蛋白水平明显下降。结论 槲皮素显著抑制肺癌干细胞活性,其机制可能与调控Wnt/β-catenin和SHH通路有关。Objective To explore the inhibitory effects of quercetin on the activity of A549 lung cancer stem cells.The role of quercetin on Wnt/β-catenin and Sonic Hedgehog( SHH) pathway was further illustrated.Method The lung cancer cell line A549 was cultured in vitro and treated with various concentrations of quercetin(0,5,10,25,50,100 and 200 μmol/L) for 24,48 and 72 h,cell vitality was detected by MTT assay.Lung cancer stem cells were isolated and enriched using sphere-forming assay.The effects of quercetin on the sphere formation and the expression of lung cancer stem cell specific markers were evaluated.The role of quercetin on the proliferation and apoptosis of sphere-forming cells was further determined by flow cytometry and western blot analysis.Finally,the protein levels of key molecules of Wnt/β-catenin and SHH pathways were detected by western blot assay.Result Our results showed that quercetin significantly inhibited A549 cell viability in a time-dependent and concertation-dependent manner.Compared to the vehicle control group,quercetin markedly suppressed the sphere formation ability of A549 cells and reduced the expression of lung cancer stem cell specific markers CD133,CD44 and aldehyde dehydrogenas 1A1( ALDH1A1).Furthermore,quercetin reduced proliferation and induced apoptosis of lung cancer stem cells.The levels of proliferating cell nuclear antigen(PCNA),Cyclin D1 and B-cell lymphoma-2( Bcl2) were markedly downregulated,while the expression of Bcl2-associated X( Bax) and cleaved cysteinyl aspartate specific proteinase( Cleaved Caspase 3) was significantly upregulated upon quercetin treatment.The Wnt/β-catenin and SHH-related proteins phospho-glucogen synthase kinase-3β( p-GSK3β)( Ser9),β-catenin,c-Myc,glioma-associated oncogene homolog 1( GLi1),GLi2 and SHH in lung cancer stem cells were also inhibited by quercetin.Conclusion Quercetin significantly suppressed the activity of lung cancer stem cells,which might be associated with the downregulation of Wnt/β-catenin and SHH pathways.

关 键 词：槲皮素 肺癌干细胞 凋亡 WNT/Β-CATENIN通路 Sonic Hedgehog通路 

分 类 号：R285[医药卫生—中药学]