miR-497抑制结肠癌细胞HCT116炎症相关基因表达的基因芯片研究

Research on miR-497 inhibiting the expression of inflammation-related genes in colon cancer cell line HCT116 using gene expression microarray

目的：采用基因芯片分析miR-497高表达对结肠癌细胞HCT116基因表达谱的影响。方法利用慢病毒感染人结肠癌细胞株 HCT116，建立 miR-497稳定高表达的结肠癌细胞模型HCT116-497细胞及阴性对照HCT116-CON细胞，用全基因组表达谱芯片筛选差异表达基因，利用MAS 3.0生物信息注释系统进行基因本体（gene ontology，GO）和信号通路富集分析（pathway analysis），筛选炎症相关基因。应用实时定量反转录聚合酶链反应（qRT-PCR）对候选基因进行验证。结果筛选出绝对倍数变化（absolute fold change）3.0倍以上的下调基因582个，发现其主要富集于炎症相关细胞因子网络等信号通路。选取下调基因中参与细胞因子网络和MAPK信号通路的15个炎症相关基因进行PCR验证后显示，与 HCT116-CON 细胞相比，HCT116-497细胞中10个基因（CACNB1、FOS、IL-29、RPS6KA2、TNFSF15、IL-11、INHBC、CSF1R、JAK3和IL-2Rβ）表达水平降低，差异均有统计学意义（均P＜0.05），与芯片结果一致。结论 miR-497抑制结肠癌细胞HCT116中炎症相关基因mRNA的表达。

Objective To analyze the effect of miR-497 high expression on the gene expression profile of colon cancer cell line HCT116. Methods MiR-497 high expressing colon cancer cell model HCT116-497 and negative control HCT116-CON were established by lentiviral transduction. The human （V2） gene expression microarray was used to identify genes that were differentially expressed between colon cancer cells overexpressing miR-497 and the controls. The candidates were subjected to the gene ontology （GO） and KEGG pathway enrichment analysis by Molecule Annotation System 3.0 （MAS3.0）. The differential expression of representative genes relative to inflammation were confirmed by real-time quantitative reverse transcription polymerase chain reaction （qRT-PCR）. Results Of all the differently expressed genes, 582 genes were down-regulated by at least 3-folds, which were enriched in inflammation-related signaling pathways in colon cancer cells overexpressing miR-497. The decrease in 15 representative genes was validated by qPCR. Compared with those in HCT116-CON cells, expressions of 10 genes in HCT116-497 cells, including CACNB1, FOS, IL-29, RPS6KA2, TNFSF15, IL-11, INHBC, CSF1R, JAK3 and IL-2Rβ, were decreased significantly, and there were statistical differences （all P〈 0.05） Conclusion MiR-497 inhibits the mRNA expression of inflammation-related genes in colon cancer cell line HCT116.