双功能基因APE1调控人骨肉瘤HOS细胞miRNAs的初步研究.docVIP

双功能基因APE1调控人骨肉瘤HOS细胞miRNAs的初步研究 戴 楠，李梦侠，卿 毅，李崇义，杨宇馨，代晓燕，王 东 （400042 重庆，第三军医大学大坪医院野战外科研究所肿瘤中心） [摘要] 目的比较APE1-konckdown骨肉瘤HOS细胞miRNAs表达谱的差异，初步探讨APE1可能调控的miRNAs及其涉及的功能通路。方法利用Ad5/F35-APE1 siRNA腺病毒敲低骨肉瘤HOS细胞中APE1的表达，用miRNA芯片检测敲组和对照组miRNAs表达谱的改变，qRT-PCR验证，利用生物信息学分析差异miRNAs涉及的功能。结果 miRNAs，APE1中有32个miRNAs表达显著改变；qRT-PCR验证个miRNAs的改变与芯片结果趋势一致其中个上调，个下调；分析发现，个显著变化的miRNAs涉及的靶基因的功能主要定位在细胞内，与蛋白、DNA结合以及转录功能有关，参与调控发育和细胞代谢等生物过程；athway分析显示其靶基因主要涉及粘附连接、轴突导向、ECM-受体相互作用、肿瘤通路，以及TGF-β、MAPK、ErbB、Wnt、mTOR及p53等关键的细胞信号通路。结论APE1能直接影响骨肉瘤细胞miRNAs的表达；生物信息学分析表明APE1通过调控miRNAs可能影响下游靶基因的表达，参与肿瘤发生、细胞生存、增殖、粘附等生物学过程和多个信号通路，与骨肉瘤的发生发展、侵袭转移密切相关。 脱嘌呤脱嘧啶核酸内切酶；；生物信息学；骨肉瘤 Effect of a double functional gene APE1 on miRNAs expression in human osteosarcoma HOS cells Dai Nan, Li Mengxia, Qing Yi, Li Chongyi, Yang Yuxin, Dai Xiaoyan, Wang Dong (Cancer Center, Institute of Surgery Research, Daping Hospital, Third Military Medical University , Chongqing, 400042, China) [Abstract] Objective To determine the differential expression profile between apurinic/apyrimidinic endonuclease (APE1) knockdown HOS cells and wild-type HOS cells, and to investigate the roles of these differential miRNAs in osteosarcoma cells. Methods After APE1 expression were down-regulated by APE1-siRNA adenovirus Ad5/F35-APE1 siRNA in HOS cells, microarray were used to confirm the change of microRNAs (miRNAs) in APE1-knockdown osteosarcoma HOS cells and wide-type cells. The results were confirmed by qRT-PCR. Comprehensive bioinformatics-based analysis, including target genes, gene ontology (GO) analysis and pathway analysis were carried out to analyze these differential expressed miRNA and their functions. Results miRNA microarray indicated that there were 32 miRNAs differentially expressed in APE1 knockdown HOS cells. qRT-PCR demonstrated that 13 miRNAs (coincidence with the results of microarray) were significantly changed ( 2-fold), with 7 of them up-regulated and the other 6 down-regulated. Furthermore, GO analysis showed that these mi