基于生物信息学途径探讨阿托伐他汀钙抗肿瘤作用的分子机制

目的基于生物信息学途径探讨与阿托伐他汀钙抗肿瘤作用相关的分子机制及生物信号通路。方法人脐静脉内皮细胞株EA.hy926分为两组,对照组加0.01%二甲基亚砜(DMSO),实验组加阿托伐他汀钙(10-5 mol/L,以0.01%DMSO溶解),共同孵育24 h,提取总RNA,用Affymetrix U133 plus 2.0全基因组表达芯片检测两组对EA.hy926细胞基因表达谱的影响。用SAM软件筛选两组之间的差异基因,应用基因富集度分析(gene set enrichment analysis,GSEA)、DAVID基因功能聚类分析软件进行通路富集分析,应用the Connectivity Map(c Map)数据库对芯片数据进行分析,并对肿瘤相关信号通路的靶基因进行RT-PCR及Westernblot验证。结果基因表达谱芯片分析显示,实验组与对照组相比,获得差异表达基因649个,其中上调基因295个,下调基因354个;GSEA富集分析提示,上调了Kruppel样转录因子等血管保护基因,下调了CCNA2、CCNE2、CCNB1和CCNB2等细胞周期相关基因,并经过RT-PCR验证。通过Cmap分析,筛选到与MS-275、trichostatin A等组蛋白去乙酰化酶抑制剂及白藜芦醇等药物有较高的相似性。结论基于生物信息学的研究发现,阿托伐他汀钙可能发挥着类似组蛋白去乙酰化酶抑制剂(HDAC inhibitors)和G1/S(开始)期及G2/M(有丝分裂)期细胞周期抑制剂的作用,为阿托伐他汀钙作为抗肿瘤药物的研究提供了可行性。

Bioinformatics-based approaches for the molecular mechanisms of the antitumour effects of atorvastatin

Objective To discover the molecular mechanism and the pathway of the anti-carcinoma effect of atorvastatin based on bioinformatics. Methods The human umbilical vein endothelial cell line EA.hy 926 was divided into two groups. The experimental group was incubated with atorvastatin (10-5 mol/L) in vitro for 24 h, and the control group was incubated with DMSO (0.01%). Total RNA from these cells was extracted and analysed for gene expression profiling. SAM microarray analysis software was used to analyse the different genes between the control and experiment groups. Gene set enrichment analysis (GSEA) and DAVID were used for gene ontology(GO) and the pathway analysis. A connectivity map(Cmap) was used to analyse the different genes. RT-PCR and Westernblot analysis were used to verify the target genes related to tumour signalling pathways. Results Compared with the control group,649 genes were detected with marked changes in the experimental group,with 295 up-regulated genes and 354 down-regulated genes in atorvastatin-treated EA.hy 926 cells. Kruppel groups in up-regulated gene expression and cell cycle genes including CCNA2,CCNE2,CCNB1 and CCNB2 in down-regulated gene expression changed with statistical significance according to GSEA and were also verified by real-time PCR. In the Cmap analysis, the profiles of the HDAC inhibitor MS-275 ,trichostatin A and resveratrol were found to be highly similar to that of atorvastatin. Conclusion Based on the bioinformatics,atorvastatin has a synergistic effect with histone deacetylase inhibitors,resveratrol and phenothiazine derivatives. Atorvastatin plays a similar role to histone deacetylase inhibitors and the cell cycle inhibitors of the G1/S phase (start) and the G2/M (mitosis) phase. These findings demonstrate the feasibility of atorvastatin as an antitumour drug.