HDAC抑制剂SB939抑制三阴性乳腺癌细胞相关基因芯片分析

Gene Microarray Analysis of HDAC Inhibitor SB939 Associated with Inhibition of Triple-negative Breast Cancer Cells

目的通过基因芯片技术分析三阴性乳腺癌细胞经SB939抑制剂作用后基因的差异表达。方法选三阴性乳腺癌细胞MDA-MB-231接种于培养皿中培养;待细胞贴壁后,用不同浓度的SB939(0、40、60μmol·L^(-1))处理24 h,将收集的细胞分为3个对照组C(未经SB939处理C1~C3)及6个实验组T(低剂量组T1~T3及高剂量组T4~T6)进行RNA的提取和纯化;随后与基因芯片杂交并进行芯片图像分析,以差异≥1.5倍的标准筛选差异表达基因。结果与对照组相比,经HDAC抑制剂SB939高剂量处理后,根据基因芯片得到的数据并分析差异表达基因,其中表达上调1.5倍的基因19个,表达下调1.5倍的基因92个;通过GSEA分析,miR-501、miR-23A、miR-23B等miRNA在表达下调基因中显著富集;通过GO富集分析发现,下调的DEGs主要富集在唾液腺形态发生发展、受体结合、蛋白结合、细胞外空隙、血小板α颗粒管腔、肌动蛋白丝等;通过KEGG通路富集分析,下调DEGs通路主要富集于上皮细胞的细菌侵袭途径,磷酸肌醇代谢、磷脂酰肌醇信号系统及ECM-受体相互作用途径。结论基因芯片技术能快速有效地检测SB939抑制剂抑制三阴性乳腺癌细胞后相关基因、miRNA基因表达水平及相关信号通路的改变,为乳腺癌精准治疗提供理论依据。

Objective The gene microarray technique was used to analyze the differential expression of genes in triple-negative breast cancer cells after the action of the SB939 inhibitor.Methods Triple-negative breast cancer cells MDA-MB-231 were selected and cultured in culture dishes.After the cells were plastered and treated with different concentrations of SB939(0,40,60μmol·L^(-1))for 24 h,the collected cells were divided into three control group C(C1 to C3 without SB939 treatment)and six experimental group T(T1 to T3 in the low dose group and T4 to T6 in the high dose group)for RNA extraction and purification.Subsequently,they were hybridized with gene microarrays,and the microarray images were analyzed,and then screened for differentially expressed genes with a difference greater than or equal to 1.5-fold.Results Compared with the control group,after high dose treatment with HDAC inhibitor SB939,according to the data obtained by gene microarray and analysis of differentially expressed genes,19 genes with 1.5-fold up-regulated expression and 92 genes with 1.5-fold down-regulated expression;analysis of miRNA by GSEA showed that miR-501,miR-23A,miR-23B and other miRNAs were significantly enriched in the down-regulated genes;the GO enrichment analysis revealed that down-regulated DEGs were mainly enriched in salivary gland morphogenesis development,receptor binding,protein binding,extracellular space,plateletαgranule lumen,actin filaments,etc;the KEGG pathway enrichment analysis showed that down-regulated DEGs pathway was mainly enriched in epithelial cells of bacterial invasion pathway,phosphatidylinositol metabolism,phosphatidylinositol signaling system,and ECM-receptor interaction pathway.Conclusion Gene microarray technology can quickly and effectively detect the changes in gene levels,miRNA levels,and related signaling pathways after the SB939 inhibitor inhibits triple-negative breast cancer cells,thus providing a theoretical basis for precise breast cancer treatment.