摘要
为探讨人红白血病HEL细胞经羊栖菜硫酸多糖(SFPSⅡ)处理后基因表达谱的变化,本研究用SFPSⅡ作用HEL细胞24 h,应用转录组技术筛选差异表达基因,分析差异基因富集的GO功能和KEGG信号通路,并通过实时荧光定量PCR(RT-qPCR)对部分差异表达基因进行验证。结果表明,SFPSⅡ处理HEL细胞后,HEL细胞活力降低,呈浓度依赖性,但对正常人胚肺成纤维细胞MRC-5几乎无毒性。与对照组相比,共有1248个基因差异表达,其中219个上调,1029个下调。GO分析显示,差异表达基因主要改变DNA构象、硫酸盐转运及对肿瘤坏死因子和外在凋亡信号反应等生物学功能。KEGG信号通路分析显示,显著富集的通路与Rap 1信号通路等癌症和免疫密切相关。试验随机选择16个显著差异基因,并通过RT-qPCR进行确认,发现SFPSⅡ诱导HEL细胞后,导致表达差异基因与肿瘤生物学进程和通路显著相关。本研究结果为揭示羊栖菜硫酸多糖抗肿瘤机制的提供了理论依据。
To study the systematic transcriptomic changes of human erythroleukemia cells(HEL) stimulated with polysaccharide fraction of Sargassum fusiforme(SFPS Ⅱ). HEL cells were treated with SFPS Ⅱ for 24 hours, the differentially expressed genes was analyzed by transcriptome technology. The genes were analyzed with Gene Oncology(GO) Term and Kyoto Encyclopedia of Gene and Genomes(KEGG) to screen the significantly enriched signals. The differently expressed genes were verified with the real-time quantitative PCR(RT-qPCR). After treated with SFPS Ⅱ, the cell proliferation of HEL cells were inhibited, cell viability was reduced in a concentration-dependent manner, while showed less toxic effect in normal human embryonic lung(MRC-5) cells. Compared with the control, a total of 1 248 differentially expressed genes(DEGs) were identified, including 219 up-regulated genes and 1 029 down-regulated genes. GO analysis indicated that differentially expressed genes mainly exercised in the molecular functions of DNA conformation change, sulfate transmembrane transport, response to tumor necrosis factor and regulation of extrinsic apoptotic signaling pathway. Pathway enrichment analysis showed that these genes involving in Rap 1 signaling pathway and immune response. Finally, with 16 DEGs which were randomly selected, and confirmed by RT-qPCR, the differentially expressed genes resulted from SFPS Ⅱ treating in HEL cells were significantly correlated with the biological processes and signaling pathways related to cancer. That could provide a theoretical basis for revealing the antitumor mechanism of sulfated polysaccharide from Sargassum fusiforme.
作者
丁浩淼
杜昊霏
付瑞洁
金旭东
谢策
陈彤妮
汪财生
钱国英
DING Haomiao;DU Haofei;FU Ruijie;JIN Xudong;XIE Ce;CHEN Tongni;WANG Caisheng;QIAN Guoying(College of Biological and Environmental Sciences,Zhejiang Wanli University,Ningbo,Zhejiang 315100)
出处
《核农学报》
CAS
CSCD
北大核心
2021年第3期595-604,共10页
Journal of Nuclear Agricultural Sciences
基金
宁波市“十三五”海洋经济创新发展示范项目资助(NBHY-2017-S5、 NBHY-2017(1))
宁波市高校食品科学与工程重点学科资助(2020FSEKDNB001)
浙江省“生物工程”一流学科资助(ZS2020011)。
关键词
羊栖菜
硫酸多糖
人红白血病
转录组
差异表达基因
Sargassum fusiforme
sulfated polysaccharide
human erythroleukemia
transcriptome
differentially expressed gene
