【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。更多还原

【Abstract】 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.更多还原