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作 者:朱连成[1] 郭骞[1] 勾睿 刘娟娟[1] 刘晴[1] 林蓓[1] ZHU Liancheng;GUO Qian;GOU Rui;LIU Juanjuan;LIU Qing;LIN Bei(Department of Obstetrics and Gynecology,Shengjing Hospital,China Medical University,Shenyang 110004,China)
机构地区:[1]中国医科大学附属盛京医院妇产科,沈阳110004
出 处:《中国医科大学学报》2020年第5期385-390,395,共7页Journal of China Medical University
基 金:国家自然科学基金(81472437,81672590);国家自然科学基金青年基金(81602438);辽宁省博士启动基金(201601133);盛京医院345人才工程计划。
摘 要:目的探讨转染人附睾蛋白4(HE4)基因对卵巢癌细胞ES-2基因表达的影响。方法构建HE4高表达及空质粒对照组细胞系,通过基因芯片探寻存在差异表达的基因,并对其进行基因本体(GO)注释和富集分析、京都基因与基因组百科全书(KEGG)通路分析,并对芯片结果进行验证。结果有639个基因发生了差异表达,其中上调基因283个,下调基因356,实时PCR及免疫组化方法对差异表达基因FOXA2和SERPIND1进行了RNA和蛋白水平的验证,结果与芯片结果相一致。GO富集分析发现差异基因在生物过程中,参与了生物高聚物代谢、程序性细胞死亡及凋亡等,KEGG通路分析发现这些基因与MAPK、性激素生物合成、癌症、细胞周期和p53信号等有关。结论与HE4相关的基因表达谱序列的变化可以为HE4在卵巢癌中功能及作用通路等分子机制的研究提供理论基础。Objective To investigate the influence of human epididymis protein 4(HE4)transfection on gene expression in the ovarian cancer cell line,ES-2.Methods Six gene chips were used to determine whether the gene expression profile differs between the HE4 transfected cell line and its empty-vector control cell line.Quantitative real-time PCR(qRT-PCR)and immunohistochemical staining were used to validate the microarray results.The differentially expressed genes(DEGs)were further applied to enrichment analysis,including the gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways.Results The gene expression profile identified 639 DEGs,in which 283 genes were upregulated and 356 genes were downregulated in response to HE4 transfection.The biological process in the GO enrichment analysis revealed that the DEGs participated in the biopolymer metabolic process,programmed cell death,and apoptosis,and these DEGs were involved in the KEGG pathways,such as MAPK,steroid biosynthesis,cancer,cell cycle,and p53 signaling.Conclusion The identification of DEGs in response to HE4 may provide a theoretical basis to evaluate its role and pathway in ovarian cancer.
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