姜黄素对胰腺癌细胞上皮间质转化的影响  

Curcumin inhibits TGF-β1-induced epithelialmesenchymal transition of pancreatic cancer cells

作　　者：庞慧芳[1,2] 覃华[1] 赵秋[1] 许琮[1] 赵慧贞[1] 朱亮[1] 黎培员[1] 李德民[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院消化内科,湖北省武汉市430030 [2]包头医学院第二附属医院消化内科,内蒙古自治区包头市014030

出　　处：《世界华人消化杂志》2014年第18期2565-2571,共7页World Chinese Journal of Digestology

摘　　要：目的:探讨姜黄素(curcumin,Cur)抑制胰腺癌细胞上皮间质转化(epithelial-mesenchymal transition,EMT)及其影响胰腺癌侵袭转移的潜在机制.方法:常规培养3种不同分化特征的胰腺癌细胞株PANC-1、AsPC-1、BxPC-3细胞,采用Western blot法检测其EMT标志物E-cadherin、Vimentin蛋白的表达.以终浓度10 ng/mL的转化生长因子-β1(transforming growth factor beta1,TGF-β1)处理低分化PANC-1细胞24 h后,在倒置显微镜下观察其形态变化.分别以10 ng/mL TGF-β1、10μmol/L Cur+10 ng/mL TGF-β1、10μmol/L Cur处理PANC-1细胞,另设空白对照组,分别采用Real-time PCR及Western blot法检测各组E-cadherin、Vimentin表达变化.结果:Western blot结果显示,PANC-1、AsPC-1、BxPC-3细胞的E-cadherin蛋白表达依次增强(1.00±00、1.36±0.04、2.14±0.06,P<0.05),而Vimentin表达则依次减弱(1.00±0.00、0.60±0.05、0.49±0.04,P<0.05),这表明低分化的PANC-1细胞间质特性最强.TGF-β1刺激PANC-1细胞发生典型的EMT形态变化.Real-time PCR和Western b l o t结果均显示,PA N C-1细胞经姜黄素及T G F-β1处理后,与对照组相比,T G F-β1组E-cadherin mRNA及蛋白表达均明显下调(分别为0.67±0.05、0.47±0.05,P<0.05 vs对照组),Vimentin基因mRNA及蛋白则明显上调(分别为2.38±0.14、1.43±0.07,P<0.05 vs对照组),说明TGF-β1促进PANC-1细胞EMT过程.而与TGF-β1组相比,Cur+TGF-β1及Cur组的E-cadherin表达具有上调趋势(mRNA分别为0.98±0.06、1.34±0.08,P<0.05 vs TGF-β1组;蛋白为0.32±0.04、0.68±0.06,P<0.05 vs TGF-β1组),Vimentin却下调(mRNA分别为0.63±0.08、0.99±0.07,P<0.05 vs TGF-β1组;蛋白分别为1.01±0.14、0.57±0.06,P<0.05vs TGF-β1组),其蛋白与mRNA表达结果基本一致.这表明姜黄素具有阻断TGF-β1诱导的PANC-1细胞EMT效应.结论:姜黄素能够阻断TGF-β1诱导的PANC-1细胞的EMT过程,从而抑制其侵袭转移.AIM： To investigate the effect of curcumin on epithelial-mesenchymal transition（EMT） of pancreatic cancer cells and the implications for pancreatic cancer invasion and metastasis. METHODS： Three pancreatic cancer cell lines PANC-1, AsPC-1 and BxPC-3, with various features of cell differentiation, were used. Total proteins extracted from them were used to determine protein expression of EMT mark-ers E-cadherin and Vimentin by Western blot. PANC-1 cells, which have the capability of low differentiation and strong invasion and metastasis, were treated with transforming growth factor beta 1（TGF-β1） at a final concentration of 10 ng/mL for 24 h, and cellular morphological changes were observed under a microscope. PANC-1 cells were then treated with 10 ng/mL TGF-β1, 10 μmol/L curcumin ＋ 10 ng/mL TGF-β1, and 10 μmol/L curcumin, respectively. Untreated PANC-1 cells were used as normal controls. Total RNA and proteins were extracted to assay the expression of E-cadherin and Vimentin by real-time PCR and Western blot. RESULTS： Western blot results showed that, the expression of E-cadherin protein in pancreatic cancer cell lines PANC-1, AsPC-1 and BxPC-3 increased in the ascending order（1.00 ± 0.00, 1.36 ± 0.04, 2.14 ± 0.06, P〈0.05）, but Vimentin decreased gradually（1.00 ± 0.00, 0.60 ± 0.05, 0.49 ± 0.04, P〈0.05）, which indicated that poorly differentiated PANC-1 cells had the most remarkable mesenchymal phenotype and stronger potential ability of invasion and metastasis than moderately and well differentiated AsPC-1 and BxPC-3 cells. TGF-β1 induced PANC-1 cells to present typical morphological changes of EMT. Real-time PCR and Western blot indicated that compared with the control group, the expression of E-cadherin in the TGF-β1 group was downregulated significantly at both mRNA and protein levels（P〈0.05）, and Vimentin expression was up-regulated remarkably（P〈0.05）, which suggested that TGF-β1 can promote the EMT of PANC-1 cells. However, compared with the TGF-β1 group,

关 键 词：姜黄素 胰腺癌 上皮间质转化 

分 类 号：R735.9[医药卫生—肿瘤]