南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (05): 606-609.

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微RNA对K562细胞基因表达谱的影响

周珏宇； 马文丽； 费嘉； 丁大鹏； 石嵘； 姜立； 郑文岭；   

  1. 南方医科大学基因工程研究所； 暨南大学医学院生物化学与分子生物学教研室； 华南基因组研究中心 广东广州510515； 广东广州510515； 广东广州510632； 广东广州510800；
  • 出版日期:2006-05-20 发布日期:2006-05-20
  • 基金资助:
    国家自然科学基金(39880032);广东省自然科学基金博士启动项目(5300488)~~

Effects of microRNA miR-181a on gene expression profiles of K562 cells

ZHOU Jue-yu1,MA Wen-li1,FEI Jia2,DING Da-peng1,SHI Rong1,JIANG Li1,ZHENG Wen-ling3 1Institute of Genetic Engineering,Southern Medical University,Guangzhou 510515,China;2Department of Biochemistry and Molecular Biology,Medical College of Jinan University,Guangzhou 510632,China;3South China Genomics Research Center,Guangzhou 510800,China   

  1. 南方医科大学基因工程研究所； 暨南大学医学院生物化学与分子生物学教研室； 华南基因组研究中心 广东广州510515； 广东广州510515； 广东广州510632； 广东广州510800；
  • Online:2006-05-20 Published:2006-05-20

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摘要： 目的应用基因芯片技术,研究微RNA(miR-181a)对人白血病K562细胞基因表达谱的影响。方法针对miR-181a成熟序列设计并合成miR-181aRNAduplexes,采用OligofectamineTM脂质体法转染K562细胞。应用AgilentHuman1A寡核苷酸基因芯片,检测和分析对照组与微RNA转染组间的基因表达谱差异。结果从20173个候选基因中,筛选出228个差异表达基因。与对照组K562细胞比较,表达上调的基因有59个,主要有代谢相关基因、抑癌基因、信号转导相关基因、免疫防御相关基因等;表达下调的有169个,主要有原癌基因、DNA结合与转录因子、代谢相关基因、信号转导相关基因、细胞周期及发育相关基因等。RT-PCR技术验证了CTCF、ZAP70、SEMA4C和RALA4个基因表达的变化。结论微RNA转染K562细胞48h后,影响了细胞内一系列基因的表达。这些基因可能与微RNA的功能相关,同时也是转录后水平上抑制基因表达的调控方式实现的基础。这为进一步深入研究微RNA的功能及其具体的作用机制提供了重要的线索和依据。 

Abstract: Objective To investigate the effect of microRNA on the gene expression profile of human leukemia K562 cells using microarray technique.Methods miR-181a RNA duplexes were designed and synthesized according to the mature sequence of miR-181a.Forty-eight hours after transfection of in vitro cultured K562 cells using OligofectamineTM,gene expression profiles of the cells were studied and analyzed using Agilent Human 1A Oligo microarray.Results Totalling 228 differentially expressed genes were identified from the 20 173 screened genes,including 59 up-regulated ones(consisting of metabolism-associated genes,tumor suppressor genes,signal transduction-associated genes,immunity and defense-associated genes etc),and 169 down-regulated ones(consisting of oncogenes,DNA-binding and transcription genes,metabolism-associated genes,signal transduction-associated genes,cell cycle and development-associated genes etc.)in the transfected K562 cells as compared with the control K562 cells.Changes in expressions of CTCF,ZAP70,SEMA4C and RALA were confirmed by semi-quantitative reverse transcription-polymerase chain reaction.Conclusions miR-181a transfection for 48 h induces gene expression profile changes in K562 cells,indicating the functionality of the miR-181a.These differentially expressed genes are related to the functions of the microRNA,and may also be the basis of the regulation model of posttranscriptional gene silencing.These findings provide an evidence for further study of the machineries and functions of the microRNA in mammalian cells. 

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