促进PPAR-γ表达对TGF-β1诱导人胰腺癌BxPc-3细胞上皮-间质转化的影响  

Effect on TGF-β1 induction of epithelial-mesenchymal transition in human pancreatic cancer BxPc-3 cells by promoting the expression of PPAR-γ

作　　者：李博[1] 施景龙 魏海雄 邱厚匡[1] 许鸣[1] LI Bo;SHI Jinglong;WEI Haixiong;QIU Houkuang;XU Ming(Department of Gastroenterology,Gangdong Second Provincial General Hospital,Guangzhou 510317;Department of General Surgery,Guangzhou 12th People’s Hospital;Shenzhen Pingshan District Center for Disease Control and Prevention,China)

机构地区：[1]广东省第二人民医院消化内科,广东广州510317 [2]广州市第十二人民医院普外科 [3]深圳市坪山区疾病预防控制中心

出　　处：《胃肠病学和肝病学杂志》2021年第2期189-193,共5页Chinese Journal of Gastroenterology and Hepatology

基　　金：广东省医学科研基金项目(A2019279)。

摘　　要：目的通过促进过氧化物酶体增殖物激活受体γ(peroxisome proliferator activated receptor-γ,PPAR-γ)的表达,分析其对TGF-β1诱导人胰腺癌BxPc-3细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)过程的影响。方法培养BxPc-3细胞,予TGF-β1诱导BxPc-3细胞,观察BxPc-3细胞形态学改变,Western blotting检测各组细胞EMT相关标志物,Transwell-matrigel实验检测细胞侵袭能力,建立BxPc-3细胞EMT模型。TGF-β1诱导前加入相应终浓度为5μmol/L、10μmol/L、20μmol/L、40μmol/L的PPAR-γ激动剂罗格列酮预处理,观察细胞侵袭能力的变化;选择20μmol/L、40μmol/L的PPAR-γ激动剂罗格列酮预处理组检测相应的PPAR-γ和EMT相关标志物上皮标记基因E-cadherin蛋白,间质标记基因Vimentin、N-cadherin蛋白的表达。结果BxPc-3细胞在TGF-β1作用下与对照组相比,其细胞形态逐渐变化为长梭形,出现EMT的特征性形态学改变;TGF-β1组穿膜细胞数较对照组差异有统计学意义(P<0.05);且TGF-β1组细胞间质标记基因Vimentin、N-cadherin蛋白表达与对照组相比均明显升高(P<0.05),上皮标记基因E-cadherin蛋白表达较对照组下降(P<0.05)。TGF-β1+20μmol/L、TGF-β1+40μmol/L罗格列酮组穿膜细胞显著低于TGF-β1组(P<0.05),而TGF-β1+5μmol/L、TGF-β1+10μmol/L罗格列酮组穿膜细胞与TGF-β1组相比,差异无统计学意义(P>0.05)。BxPc-3细胞经TGF-β1作用后PPAR-γ表达较对照组明显下降(P<0.05),而TGF-β1+20μmol/L、TGF-β1+40μmol/L罗格列酮组细胞PPAR-γ表达逐渐升高(P<0.05);TGF-β1+20μmol/L、TGF-β1+40μmol/L罗格列酮组细胞上皮标记基因E-cadherin蛋白表达较TGF-β1组逐渐升高(P<0.05);TGF-β1+20μmol/L、TGF-β1+40μmol/L罗格列酮组细胞间质标记基因Vimentin、N-cadherin蛋白表达与TGF-β1组相比均明显降低(P<0.05),而前两组比较,差异无统计学意义(P>0.05)。结论TGF-β1可以诱导人胰腺癌BxPc-3细胞EMT;促进PPAR-γ表达可以有效抑制人胰腺癌BxPObjective To investigate the effect on transforming growth factor beta-1(TGF-β1)induction of epithelial-mesenchymal transition(EMT)in human pancreatic cancer BxPc-3 cells by promoting the expression of peroxisome proliferator activated receptor-γ(PPAR-γ).Methods TGF-β1 was used to induce EMT in BxPc-3 cells.The morphological changes were observed under phase-contrast microscope.The changes of EMT-related markers in BxPc-3 were analyzed by Western blotting.Direct invasive test was determined by Transwell-matrigel invasion chambers.BxPc-3 was pretreated by PPAR-γagonist Rosiglitazone with a final concentration of 5μmol/L,10μmol/L,20μmol/L and 40μmol/L before induction with TGF-β1,the change of cell invasion ability were observed.The expressions of PPAR-γand EMT-related markers E-cadherin,Vimentin,N-cadherin with the Rosiglitazone pretreatment group of 20μmol/L and 40μmol/L were detected.Results The BxPc-3 cells became elongated after exposed to TGF-β1,characteristic morphological changes of EMT were observed.The number of membrane penetrating cells in TGF-β1 group was significantly different from that in control group(P<0.05).Expressions of plasma marker genes Vimentin and N-cadherin in TGF-β1 group was significantly higher than that in the control group(P<0.05).The expression of E-cadherin protein decreased compared with the control group(P<0.05).The number of membrane penetrating cells in TGF-β1+20μmol/L,TGF-β1+40μmol/L Rosiglitazone group was significantly lower than that in TGF-β1 group(P<0.05).There was no significant difference between TGF-β1+5μmol/L,TGF-β1+10μmol/L Rosiglitazone group and TGF-β1 group(P>0.05).PPAR-γexpression of BxPc-3 cells was significantly lower after treatment with TGF-β1(P<0.05),PPAR-γexpression gradual increase in TGF-β1+20μmol/L,TGF-β1+40μmol/L Rosiglitazone group(P<0.05).The expression of E-cadherin in TGF-β1+20μmol/L,TGF-β1+40μmol/L Rosiglitazone group was higher than that in TGF-β1 group(P<0.05).The expressions of Vimentin and N-cadherin in TGF

关 键 词：过氧化物酶体增殖物激活受体Γ 胰腺癌 上皮-间质转化 

分 类 号：R735.9[医药卫生—肿瘤]