月腺大戟素A抗乳腺癌作用机制转录组分析  

Anti-breast cancer mechanism of transcriptome analysis of ebracteolatain A

作　　者：覃福礼 赵亮 周艳卿 周瑾 须秋萍 李群英 焦杨[1] QIN Fu-li;ZHAO Liang;ZHOU Yan-qing;ZHOU Jin;XU Qiu-ping;LI Qun-ying;JIAO Yang(Department of Pharmacology,School of Pharmacy,Guangxi Medical University,Nanning 530021,China;Department of Pharmacy,Luodian Hospital of Baoshan District of Shanghai,Shanghai 201908,China)

机构地区：[1]广西医科大学药学院药理教研室,广西南宁530021 [2]上海市宝山区罗店医院药剂科,上海201908

出　　处：《中草药》2020年第4期1003-1015,共13页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81303300);上海宝山区科技创新基金(19-E-2)。

摘　　要：目的利用二代测序技术筛选出具有显著性变化的基因,并探讨月腺大戟素A在转录组学层面上抗乳腺癌活性的作用机制。方法从月腺大戟药材中提取乙酰间苯三酚类化合物月腺大戟素A,对MCF-7细胞(乳腺癌细胞的luminal A型)进行干扰,观察被干扰后的细胞与正常细胞差异基因表达,采用二代高通量测序平台(IlluminaHi-Seq)测序技术分别对对照组和实验组各3个样本进行高通量转录组测序并进行数据分析。结果对照组和实验组分别总获得123656848、123974262个干净序列(clean reads),分别对比到参考基因组上的序列为119762214、119881622,各占总数的96.85%、96.69%;2组转录组对照可得:差异基因总数为1695个,其中上调基因770个,下调基因925个,可清楚注释的基因有3874个。应用基因本体论(gene ontology,GO)和京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)进行生物功能富集分析,GO分析发现这3874个基因主要涉及生物过程(1270个)、细胞组成(1322个)与分子功能(1282个)3个大类的45个小类,包括细胞的生长发育过程、信号蛋白活性、膜以及基因表达的调控等过程;KEGG分析发现差异表达基因涉及263条信号通路,主要代谢通路为PI3K-Akt信号通路、MAPK信号通路;以及碳水化合物代谢、心肌系统和细胞生殖系统等生物过程。结论利用二代高通量测序平台测序技术一共筛选、鉴定出差异基因1695个,更深入了解了月腺大戟素A与MCF-7细胞基因之间的相互关系,为乳腺癌治疗提供了一些理论基础。Objective To research the mechanism of ebracteolatain A against breast cancer cells, screening the genes with significant changes using second-generation sequencing, and explore the anti-breast cancer mechanism of action of ebracteolatain A at the transcriptomics level. Methods The acetyl phloroglucinol compound ebracteolatain A was extracted from Euphorbia ebracteolata, interferencing with MCF7 cells(luminal A type of breast cancer cells) to observe differential gene expression between the interfered cells and normal cells. High-throughput transcriptome sequencing and data analysis were performed on three groups of control groups and three experimental groups using Illumina Hi-Seq sequencing technology. Results A total of 123 656 848, 123 974 262 available reads were obtained in the control group and experimental group, respectively, the reads on the reference genome were 119 762 214, 119 881 622, respectively, accounting for 96.85% and 96.69% of the total;Two groups of transcriptome controls were available: the total number of differential genes was 1 695, of which 770 were up-regulated, 925 were down-regulated, and 3 874 genes were clearly annotated. Bio-enrichment analysis was carried out by using Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG). GO analysis found that these 3 874 genes mainly involved in biological processes(1 270), cell composition(1 322) and molecular function(1 282), 45 subcategories of three major categories, including cell growth and development, signaling protein activity, membrane and regulation of gene expression. KEGG analysis revealed that the differentially expressed genes involved 263 signaling pathways;The main metabolic pathways were: PI3 K-Akt signaling pathway, MAPK signaling pathway, carbohydrate metabolism, myocardial system and cellular reproductive system and etc. Conclusion The results showed that 1 695 differential genes were screened and identified by Illumina Hi-Seq sequencing technology, and the relationship between the genes of ebracteolatain A an

关 键 词：月腺大戟素A 乳腺癌 RNASEQ 转录组 功能基因 代谢通路 高通量测序平台 

分 类 号：R285.5[医药卫生—中药学]