槲皮素联合si-FSCN1对胃癌MKN45细胞增殖与侵袭和凋亡的影响  

Effect of quercetin combined with si-FSCN1 on proliferation,invasion and apoptosis of gastric cancer MKN45 cells

作　　者：张灿灿 谭志军[1] 徐敏 马志承[2] 姜伟[2] ZHANG Can-can;TAN Zhi-jun;XU Min;MA Zhi-cheng;JIANG Wei(I.Tianjin Medical University,Central Clinical College,Tianjin 300070,China;Department of General Surgery,Tianjin First Central Hospital,Tianjin 300384,China)

机构地区：[1]天津医科大学中心临床学院,天津300070 [2]天津市第一中心医院普外科,天津300384

出　　处：《中国临床药理学杂志》2020年第9期1132-1135,共4页The Chinese Journal of Clinical Pharmacology

摘　　要：目的探讨槲皮素联合si-FSCN1对胃癌MKN45细胞增殖、侵袭和凋亡的影响及其机制。方法以0、10,20,40,80和160μmol·L^-1槲皮素作用于体外培养的MKN45细胞24 h后,采用噻唑蓝(MTT)法检测细胞存活率并计算出槲皮素的半抑制浓度。将si-FSCN1和si-NC分别转染至MKN45细胞,分别命名为si-FSCN1组和si-NC组,未经转染的细胞作为空白组,采用逆转录-聚合酶链反应和蛋白质印迹法检测转染效果。以80μmol·L^-1槲皮素处理转染si-FSCN1和si-NC的MKN45细胞,分别命名为槲皮素+si-FSCN1组和槲皮素+si-NC组,用噻唑蓝和克隆形成实验检测si-FSCN1组、si-NC组、槲皮素+si-FSCN1组和槲皮素+si-NC组细胞增殖,用Transwell小室实验检测细胞侵袭,用膜联蛋白V-FITC(Annexin V-FITC)/碘化丙啶(PI)双染法检测细胞凋亡,用Western blot检测细胞中无翅基因(Wnt)/β-连环蛋白(β-catenin)信号通路相关蛋白β-catenin、细胞周期蛋白D1(Cyclin D1)、基质金属蛋白酶9(MMP-9)、血管内皮细胞生长因子(VEGF)、生存素(Survivin)蛋白的表达情况。结果槲皮素的半抑制浓度为85.60μmol·L^-1。si-NC组、si-FSCN1组、槲皮素+si-NC和槲皮素+si-FSCN1组细胞的存活率分别为(100.00±10.11)%,(51.72±3.28)%,(66.55±3.46)%和(38.95±2.35)%;克隆形成率分别为(36.48±4.05)%,(21.62±2.36)%,(28.81±2.42)%和(15.54±1.38)%,细胞凋亡率分别为(3.16±1.55)%,(20.28±3.05)%,(15.87±2.32)%和(33.45±3.28)%;侵袭细胞数分别为(88.85±6.28),(55.00±3.25),(62.68±4.52)和(36.76±3.02)个;β-catenin蛋白相对表达量分别为0.76±0.05,0.42±0.03,0.55±0.04和0.26±0.02;Cyclin D1蛋白相对表达量分别为1.21±0.22,0.72±0.06,0.84±0.06和0.32±0.03;MMP-9蛋白相对表达量分别为0.95±0.07,0.66±0.04,0.80±0.05和0.41±0.03;VEGF蛋白相对表达量分别为1.42±0.25,0.84±0.06,0.98±0.06和0.67±0.04;Survivin蛋白相对表达量分别为1.33±0.17,0.90±0.08,1.10±0.06和0.58±0.04。si-FSCN1、槲皮素+si-NC和槲皮素+si-FSCN1组分�Objective To investigate the effects of quercetin combined with si-FSCN1 on proliferation,invasion and apoptosis of gastric cancer MKN45 cells and its mechanism.Methods MKN45 cells cultured in vitro were treated with 0,10,20,40,80 and 160μmol·L-1 quercetin for 24 h.Cell viability was measured by methylthiazolyldiphenyl-tetrazolium bromide(MTT)assay and the semi-inhibitory concentration of quercetin was calculated.si-FSCN1 and si-NC were transfected into MKN45 cells,respectively,named si-FSCN1 group and si-NC group,and untransfected cells were used as blank group,and the transfection effects were detected by reverse transcription-polymerase chain reaction(RT-PCR)and Western blot.MKN45 cells transfected with si-FSCN1 and si-NC were treated with 80μmol·L^-1 quercetin,named quercetin+si-FSCN1 group and quercetin+si-NC group,cell proliferation of the si-FSCN1 group,si-NC group,quercetin+si-FSCN1 group and quercetin+si-NC group were measured by MTT and cloning formation experiment,cell invasion was detected by Transwell chamber assay,and the apoptosis was detected by double staining with Annexin V-FITC/PI,the wingless(Wnt)/β-catenin signaling pathway-related proteinsβ-catenin,Cyclin D1,MMP-9,vascular endothelial growth factor(VEGF),survivin(Survivin)protein expression were detected by Western blot.Results The semi-inhibitory concentration of quercetin was 85.60μmol·L^-1.The survival rates of cells in si-NC group,si-FSCN1 group,quercetin+si-NC group and quercetin+si-FSCN1 group were(100.00±10.11)%,(51.72±3.28)%,(66.55±3.46)%and(38.95±2.35)%,respectively.The clone formation rates were(36.48±4.05)%,(21.62±2.36)%,(28.81±2.42)%and(15.54±1.38)%,respectively.The number of invasive cells was(88.85±6.28),(55.00±3.25),(62.68±4.52)and(36.76±3.02),respectively.The apoptotic rates were(3.16±1.55)%,20.28±3.05)%,15.87±2.32)%and(33.45±3.28)%,respectively.The expression levels ofβ-catenin protein were 0.76±0.05,0.42±0.03,0.55±0.04 and 0.26±0.02,respectively.The expression levels of CyclinD1 protein were 1.2

关 键 词：胃癌 槲皮素 FSCN1 细胞增殖 细胞侵袭 细胞凋亡 无翅基因/β-连环蛋白信号通路 

分 类 号：R97[医药卫生—药品]