基于表达谱芯片的冬凌草甲素抑制胰腺癌BxPC-3细胞的机制研究  

Study on The Inhibitory Mechanism of Oridonin in Pancreatic Cancers BxPC-3 Cells by DNA Microarray

作　　者：岳静[1] 沈雯[1] 许健[1] 牟一平[2] 张婷[1] 张斌[1] 

机构地区：[1]浙江中医药大学,杭州310053 [2]浙江大学附属邵逸夫医院

出　　处：《浙江中医药大学学报》2013年第5期606-612,共7页Journal of Zhejiang Chinese Medical University

基　　金：浙江省自然科学基金(LY12H16007;Y2080506);浙江省教育厅一般项目(Y201121464);浙江省大学生科技创新活动计划暨新苗人才计划项目(2012R410015)~~

摘　　要：[目的]应用表达谱芯片比较冬凌草甲素干预前后人胰腺癌细胞株BxPC-3的基因表达差异,寻找细胞增殖和凋亡的调控分子和信号通路,探讨冬凌草甲素抗肿瘤的分子机制。[方法]提取空白对照组及冬凌草甲素作用组总RNA,并利用标准的变性琼脂糖凝胶电泳检测其完整性。借助安捷伦快速扩增标记试剂盒标记样品,与基因表达谱芯片于安捷伦杂交室进行杂交,安捷伦DNA微阵列扫描仪扫描芯片收集数据,经分析得出药物作用后差异表达的基因。[结果]32μg·mL-1冬凌草甲素组与空白对照组相比,显著上调基因有520种,显著下调基因有651种,其中Caspase-3、Caspase-7、Caspase-8、Caspase-9等基因和蛋白经PCR和western blot得到验证。[结论]冬凌草甲素对胰腺癌细胞BxPC-3基因表达谱有一定影响,其抗肿瘤和免疫调节作用可能与一系列肿瘤及免疫相关基因的表达变化有关。[Objective]To find out the regulatory molecules in the signal pathways on cell proliferation or apoptosis,and explore the inhibitory mechanism of oridonin on pancreatic tumor,we compared the difference of genes expression between treatment with and without oridonin in pancreatic cancer BxPC-3 cells.[Method]Total RNA was extracted using Trizol after treated by oridonin 32μg/ml for 36 hours,and the integrity and concentration of total RNA were assessed by electrophoresis on a denaturing agarose gel.Agilent Quick Amp Labeling Kit was used for sample labeling.Hybridization was performed in Agilent’s SureHyb Hybridization Chambers.Slides were scanned with the Agilent DNA microarray Scanner.The differentially expressed genes were screened after data analysis.[Results]Compared with two groups,520 genes up-regulated and 651 genes down-regulated were screened.Moreover,the expressions of Caspase-3,Caspase-7,Caspase-8,Caspase-9,etc.were verified by PCR and western blot.[Conclusion ]Some genes expression was inhibited by oridonin in pancreatic cancer BxPC-3 cells.Series changes of the gene expression concerning tumor and immunity might be related to the anti-tumor and immunomodulatory effect of oridonin.

关 键 词：基因芯片 冬凌草甲素 胰腺癌 BXPC-3 

分 类 号：R282.71[医药卫生—中药学]