白藜芦醇对胰腺癌细胞Panc-1的抑制作用及机制探讨  

The inhibitory effect of resveratrol on the growth of pancreatic cancer cell line Panc-1 and its related mechanism

作　　者：莫文辉 郭传勇[2] 田怡[1] 万红宇[1] 张晓红[1] 刘建生[1] 

机构地区：[1]上海瑞金医院集团闵行区中心医院消化内科,上海201199 [2]同济大学附属上海市第十人民医院肝胆胰内科,上海200072

出　　处：《肿瘤》2014年第8期685-690,共6页Tumor

摘　　要：目的：探讨白藜芦醇（resveratrol,Res）对胰腺癌细胞Panc-1的抑制作用及可能的作用机制。方法：采用不同浓度Res处理胰腺癌Panc-1细胞后,CCK-8（cell counting kit-8）法检测细胞增殖抑制率,FCM法检测细胞周期分布,实时荧光定量PCR法检测Hedgehog信号通路成员胶质瘤相关癌基因1（glioma-associated oncogene1,Gli-1）基因表达的变化。Annexin-PI双染法检测100μmol/L Res作用24 h后Panc-1细胞的凋亡情况。以针对Gli-1基因的短发夹RNA-慢病毒感染建立的稳定低表达Gli-1的Panc-1细胞为阳性对照组,12.5μmol/L Res处理的Panc-1细胞为实验组,同时设未处理及空病毒载体感染的Panc-1细胞为空白及阴性对照组,采用实时荧光定量PCR法检测各组细胞中Gli-1及其下游靶基因Patched、cyclin D1和Bcl-2的表达水平。结果：Res对Panc-1细胞生长具有抑制作用,并呈时间和浓度依赖性。随着作用浓度（12.5～100μmol/L）增加,Res对Panc-1细胞呈现出G0/G1期细胞周期阻滞作用。100μmol/L Res作用Panc-1细胞24 h后,细胞早期凋亡率为（19.1±1.25）%,显著高于未处理对照组的（2.3±0.26）%（P〈0.05）。小剂量（12.5μmol/L）Res即可对Hedgehog信号通路成员的表达产生抑制作用,与空白及阴性对照组相比,Res处理实验组及Gli-1低表达阳性对照组中Gli-1、Patched、Bcl-2和cyclin D1 mRNA表达均下调（P〈0.05）。结论：Res可抑制胰腺癌Panc-1细胞增殖,其作用机制可能与调控Hedgehog/Gli-1信号通路有关。Objective： To investigate the inhibitory effect of resveratrol （Res） on the growth of human pancreatic cancer Panc-1 cells in vitro, and explore its possible mechanism. Methods： The cell counting kit-8 （CCK-8） assay was used to detect the proliferation of Panc-1 cells after treatment by defferent concentrations of Res. After treatment with 100 μmol/L Res for 24 h, the apoptosis and cell cycle distribution were examined by Annexin-PI double staining method and flow cytometry, respectively. The expression of glioma-associated oncogene 1 （Gli-1） gene, a member of Hedgehog signal pathway, was detected by real-time fluorescent quantitative-PCR （RFQ-PCR）. Then the Panc-1 cells were transfected with short hairpin RNA （shRNA）-Gli-1 lentivirus （as the positive control） or blank lentivirus （as the negative control）, and treated with 12.5 μmol/L Res （as the experimental group） and untreated with any interventions （as the blank control）, respectively. The expressions of Gli-1 and its related downstream target genes including Patched, cyclin D1 and Bcl-2 in each group were detected by RFQ-PCR. Results： Res inhibited the growth of Panc-1 cells in a dose- and time-dependent manner in vitro. After treatment with 12.5-100 μmol/L Res for 24 h, the percentage of Panc-1 cells in G0/G1 phase was significantly higher than that of the untreated group （P 〈 0.05）. Apoptosis rate of Panc-1 cell treated with 100 μmol/L Res for 24 h was （19.1±1.25）%, which was significantly higher than that of the untreated group [（2.3±0.26）%, P 〈 0.05]. As compared with the blank and negative control groups, the expressions of Gli-1, Patched, cyclin D1 and Bcl-2 mRNAs were down-regulated in 12.5 μmol/L Res treatment group and shRNA-Gli-1transfection group （P 〈 0.05）. Conclusion： Res can suppress the proliferation of human pancreatic cancer Panc-1 cells, which may be related to the regulation of Hedgehog-Gli-1 signaling pathway.

关 键 词：胰腺肿瘤 抗肿瘤药(中药) 信号转导 刺猬蛋白质类 白藜芦醇 核转录因子Gli-1 

分 类 号：R735.9[医药卫生—肿瘤]