丹参酮ⅡA促进Eca109细胞凋亡转录组学研究

Transcriptomics study on Tanshinone ⅡA promoting apoptotic effect in esophageal squamous carcinoma Eca109 cells

目的利用RNA测序(RNA sequencing,RNA-seq)进行丹参酮ⅡA促食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)Eca109细胞凋亡的转录组学研究,探讨可能的分子机制。方法丹参酮ⅡA(8μg/ml)处理Eca109细胞48 h,空白组作为对照,进行高通量转录组测序分析,筛选差异基因,分析转录组表达谱,利用GO(gene ontology)功能注释和KEGG(kyoto encyclopedia of genes and genomes)通路富集方法探讨丹参酮ⅡA促进Eca109细胞凋亡分子机制。结果转录组测序分别获得包含28,667,820、27,830,388条过滤后测序读数,构建测序文库比对分析显示,差异基因共有627条,其中7条基因表达上调,620条基因表达下调,483条基因存在不同类型可变剪接。GO分类及KEGG通路分析表明:差异表达基因涉及多种食管癌肿瘤信号通路NF-κB、Wnt、Hedgehog,且与ESCC发生发展的进程相关。结论丹参酮ⅡA促进Eca109细胞凋亡分子机制可能与下调肿瘤信号通道相关,也进一步为ESCC治疗提供新依据。

Objective To analysis transcriptome profile and investigate the associated molecular mechanism on Tanshinone ⅡA promoting apoptotic effect by RNA-Seq in esophageal squamous carcinoma Eca109 cells.Methods The Eca109 cells were treated with Tan ⅡA（8μg/m L） for 48 hours and blank group served as control.RNA-Seq was performed and the sequencing data were compared with the reference genome.Then the differential expression profile of genes were screened,identified and analyzed in Eca109 cells.Gene ontology（GO） and kyoto encyclopedia of genes and genomes（KEGG） were analyzed accordingly.Results In brief,transcriptomic sequencing data showed 28,667,820 and 27,830,388 normalized reads were aligned and mapped respectively.Gene set enrichment analysis suggested that 627 differential gene expression profiles were clearly distinguished.Among them,7 genes were upregulated and 620 genes were downregulated genes and 483 variable splicing patterns were identified.Through GO analysis,we found that these were associated with developmental process and multicellular organismal development.Furthermore,KEGG analysis showed that some of the identified signalling pathways,such as NF-κB,Wnt,Hedgehog（Hh）,were involved with esophageal squamous cell carcinoma（ESCC） development and progression.Conclusion Tanshinone ⅡA has been shown to promote apoptosis and associated with downregulation of the Hh and other signaling pathway,then new tools for ESCC treatment may be provided further.