miR-519d过表达促使宫颈癌SiHa细胞基因表达谱发生改变

Over-expression of miR-519d alters gene expression profiles of cervical cancer SiHa cells

目的分析miR-519d转染前后人宫颈癌SiHa细胞基因表达谱的变化。方法向人宫颈癌SiHa细胞中转染模拟物上调细胞内miR-519d的表达,采用人全基因组表达谱芯片筛选其对细胞基因表达谱的影响,结合生物信息学软件进行靶基因预测,确定miR-519d的候选靶基因,采用qRT-PCR进行验证。结果 miR-519d表达上调后,共筛选到差异表达基因5172个,其中上调基因2476个,下调基因2796个。结合生物信息学软件进行靶基因预测,共确定164个miR-519d的候选靶基因。随后,对上述基因进行KEGG信号通路分析,采用STRING和pSTIING在线数据库构建候选基因编码蛋白的互作网络、预测转录相关和物理相关关系,发现了一些重要的节点分子。最后,采用qRT-PCR结果对部分关键基因进行了验证,与芯片结果基本一致。结论miR-519d过表达促使人宫颈癌SiHa细胞的基因表达谱发生了显著地改变,结合生物信息学软件获得了164个候选靶基因,为系统、全面地阐明miR-519d在宫颈癌中的作用及其分子机制提供了重要的实验依据。

Objective To investigate the alterations in gene expression profiles of cervical cancer cell line SiHa over-expressing miR-519 d. Methods SiHa cells were transfected with a miR-519 d mimic or a negative control（NC） and the changes in gene expression profiles were examined using Nimble Gen human gene expression microarray. Bioinformatics approaches based on the microarray data were used to identify the targeted genes of miR-519 d. Real-time quantitative PCR was employed to confirm the expression of the potential target genes. Results A total of 5172 genes were found to be differentially expressed in SiHa cells over-expressing miR-519 d, including 2476 up-regulated and 2796 down-regulated genes. We identified 164 potential target genes of miR-519 d, and their functions were predicted using Gene Ontology（GO） and Kyoto Encyclopedia of Genes and Genomes（KEGG） biological pathway analysis, and STRING database and pSTIING were used to search the key nodes in the protein-protein interactions and transcriptional regulatory networks in cancer. Real-time quantitative PCR confirmed the differential expressions of several candidate target genes. Conclusion Over-expression of miR-519 d alters gene expression profiles in SiHa cells. The 164 target genes of miR-519 d we identified may provide insights into the role of miR-519 d in cervical tumorigenesis.