冬凌草甲素抗食管鳞癌转录的组学研究  

Genomic study on oridonin against transcriptome of esophageal squamous cell carcinoma

作　　者：王伟[1] 丁爽 龚美源 李娜娜 燕迪迪 倪凯园 郭建成 张旗[2,3] WANG Wei;DING Shuang;GONG Meiyuan;LI Nana;YAN Didi;NI Kaiyuan;GUO Jiancheng;ZHANG Qi(Department of Oncology,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;School of Pharmaceutical Sciences,Zhengzhou University,Zhengzhou 450001;State Key Laboratory of Esophageal Cancer Prevention&Treatment,Zhengzhou 450052;Center for Precision Medicine,Zhengzhou University,Zhengzhou 450052)

机构地区：[1]郑州大学第一附属医院肿瘤科,郑州450052 [2]郑州大学药学院,郑州450001 [3]省部共建食管癌防治国家重点实验室,郑州450052 [4]郑州大学精准医学中心,郑州450052

出　　处：《郑州大学学报（医学版）》2021年第4期464-470,共7页Journal of Zhengzhou University(Medical Sciences)

基　　金：河南省科技攻关计划项目(182102310094,162102310153,152102310180,142102310330);河南省自然科学基金项目(162300410291)。

摘　　要：目的:探索冬凌草甲素对食管鳞癌细胞增殖和凋亡的影响及其作用的组学研究。方法:取食管鳞癌细胞系EC109和TE1细胞,分别用0、10、20、40、80和160μmol/L的冬凌草甲素处理,采用MTT法和流式细胞仪检测冬凌草甲素对EC109和TE1细胞增殖及凋亡的影响;冬凌草甲素处理食管鳞癌细胞后提取总RNA,采用全转录组分析技术(RNA-seq)探索食管鳞癌细胞转录组的变化;运用GO聚类分析对差异表达的基因进行功能分析。结果:冬凌草甲素可抑制食管鳞癌EC109和TE1细胞增殖,促进细胞凋亡(P<0.05);RNA-seq分析结果显示,在EC109细胞中冬凌草甲素可使463个基因表达上调,419个基因下调,在TE1细胞中冬凌草甲素可使2023个基因表达上调,1642个基因下调;其中,共同上调的基因284个,共同下调的基因176个。冬凌草甲素处理后细胞出现明显的热激反应,HSPA1A、HSPA1B、BAG3、HSPH1、DNAJB1、HSPA8、HSP90AA1、DNAJA4和DNAJA1等基因表达升高,同时作为调节GSH-ROS的TRIM16、SLC7A11、TXNRD1、SRXN1、GCLM和GCLC等的基因表达也升高。GO聚类分析显示,差异表达基因为热激反应基因、蛋白错误折叠相关基因和神经投射发生基因。结论:冬凌草甲素抑制食管鳞癌细胞的增殖并促进其凋亡,其分子机制与热激反应及GSH-ROS系统密切相关。Aim:To investigate the effect of oridonin on the proliferation and apoptosis of esophageal squamous cell carcinoma(ESCC)cell lines and its molecular mechanism of anti-esophageal cancer.Methods:ESCC cell lines EC109 and TE1 were divided into the experimental group and control group,the experimental group was treated with 0,10,20,40,80 and 160μmol/L oridonin,and the ESCC cells treated with DMSO as the control group.MTT assay and flow cytometry were respectively used to detect the effects of oridonin on the proliferation and apoptosis of EC109 and TE1 cells.Total RNA was extracted from ESCC cell lines treated with oridonin,and the transcriptome changes of ESCC cells were analyzed by RNA-seq technique.Functional analysis of differentially expressed genes was carried out through GO.Results:The proliferation inhibition rates of EC109 and TE1 cells increased after treatment with different concentrations of oridonin compared with the control group(P<0.05)and the apoptotic rate of the treatment groups was increased(P<0.05).The results of RNA-seq showed that after oridonin treatment 463 genes was up-regulated and 419 genes were down-regulated in EC109 cells and 2023 genes was up-regulated and 1642 genes were down-regulated in TE1 cells,we also found that 284 genes were up-regulated and 176 genes were down-regulated both in EC109 and TE1 cells.The expressions of heat shock genes such as HSPA1A,HSPA1B,BAG3,HSPH1,DNAJB1,HSPA8,HSP90AA1,DNAJA4 and DNAJA1 significantly increased.Genes regulating GSH-ROS such as TRIM16,SLC7A11,TXNRD1,SRXN1,GCLM and GCLC were also up-regulated.GO analysis showed that the differentially expressed genes were heat shock response genes,protein misfolding related genes and neuroprojection generating genes.Conclusion:Oridonin can inhibit the proliferation and promote the apoptosis of esophageal carcinoma cells and the anti-tumor mechanism of oridonin is closely related to the heat shock response and the GSH-ROS system.

关 键 词：冬凌草甲素 食管鳞癌 RNA-SEQ GSH-ROS系统 细胞热激反应 

分 类 号：R735[医药卫生—肿瘤]