miRNA-497与miRNA-195基因簇在宫颈癌组织中的表达及预测靶基因的生物信息学分析

The expression of miRNA-497 and miRNA-195 cluster in cervical cancer tissues and bioinformatics analysis of predicted target genes

目的探讨miRNA-497-195基因簇在宫颈癌组织中的表达,并对其预测的靶基因进行生物信息学分析,为miRNA-497-195基因簇在宫颈癌发生中的作用机制研究提供理论基础。方法收集2010年3月～2012年3月在广西壮族自治区人民医院就诊患者的宫颈标本,利用miRNA芯片技术检测宫颈病变组织中miRNA表达谱,用real-time RT-PCR进行验证;采用real-time RT-PCR检测miRNA-497和miRNA-195在宫颈病变组织中的表达情况,并用Spearman法进行miRNA-497与miRNA-195相关性分析。通过生物信息学预测miRNA-497和miRNA-195的靶基因,并对其靶基因进行GO（gene ontology）功能富集分析及信号转导通路富集分析。结果芯片结果显示,宫颈癌及高度鳞状上皮内瘤变中miRNA-195和miRNA-497表现为下调（均P〈0.05）。real-time RT-PCR结果显示,miRNA-497和miRNA-195在宫颈癌组织中表现为下调,且表达存在显著性相关（r=0.983,P〈0.05）。miRNA-497和miRNA-195预测靶基因大量重合,且集合功能也大量重合,并富集于细胞周期调控、生物学过程调控、转录调控、基因表达及核苷酸代谢过程等生物学过程,以及蛋白结合、核酸结合、蛋白激酶活性、连接酶活性等分子功能（P〈0.01）;KEGG通路分析涉及癌症通路、p53信号通路、GnRH信号通路、细胞外因子信号通路等信号传导通路及直肠癌、前列腺癌、甲状腺癌、黑色素瘤、慢性粒细胞性白血病、非小细胞肺癌等疾病通路（P〈0.05）。结论 miRNA-195和miRNA-497在宫颈癌组织中呈现低表达,二者表达呈显著正相关,miRNA-497-195基因簇可能是宫颈癌的抑制因子。miRNA-497和miRNA-195预测靶基因功能大量重合,并显著富集在与肿瘤发生相关的信号通路中。

Objective To investigate the expression of miRNA-497-195 cluster in cervical cancer tissues, and bioinformatically analyze the target genes of miRNA-497-195 cluster, in order to provide theoretical basis for further studies of miRNA-497-195 cluster biological function in cervical cancer. Methods From March 2010 to March 2012,cervical specimens were collected from the People＇s Hospital of Guangxi Zhuang Autonomous Region. miRNA array was applied to detect miRNA expression profile in cervical lesion tissues, and real-time RT-PCR was used for validation. The relative expression level of miRNA-497 and miRNA-195 were analyzed by real-time RT-PCR, and Spearman was used for the correlation analysis. The bioinformatic analysis of the target genes of miRNA-497 and miRNA-195 involved gene ontology（GO） and signal transduction pathway enrichment. Results miRNA-497 and miRNA-195 expression decreased in cervical cancer tissues compared with normal cervical tissues（P 〈0.05）. A significant positive correlation between the expression levels of miRNA-195 and miRNA-497 in the clinical samples were revealed（r = 0.983, P 〈0.05）. The predicted target genes of miRNA-497 and miRNA-195 were a lot of overlap. The functions of predicted target genes of miRNA-497 and miRNA-195 were enriched in cell cycle, regulation of biological process, regulation of transcription,regulation of gene expression, regulation of the nucleic acid metabolic process, protein binding, nucleotide binding, kinase activity, ligase activity and other biological processes and molecular functions（P 〈0.01）. In KEGG pathway, the predicted target genes involved in pathway in cancer, p53 signaling pathway, GnRH signaling pathway, Wnt signaling pathway, colorectal cancer, prostate cancer, thyroid cancer, melanoma, chronic myeloid leukemia, non-small cell lung cancer, and other signal transduction pathways and human diseases pathway（P 〈0.05）. Conclusion miRNA-497 and miRNA-195 expression decreases and obviously correlative in cervical cancer tissues,suggesting that miRNA-497-195 cluster may function as tumor suppressors in cervical cancer. Some of the predicted target genes of miRNA-497 and miRNA-195 are significantly enriched in tumor related with signaling pathways.