蜂胶黄酮PB3A作用下结肠癌细胞株中miRNA-198和miRNA-296-5p的表达及功能预测

发布时间：2018-06-05 16:50

本文选题：短叶松素--乙酸脂 + 结肠癌SW细胞　；参考：《中国病理生理杂志》2017年07期

【摘要】：目的:探讨蜂胶黄酮短叶松素-3-乙酸酯(pinobanksin-3-acetate,PB3A)对结肠癌SW480细胞微小RNA(microRNA,miRNA)表达谱的影响,为结肠癌的治疗及靶向药物研发提供理论依据。方法:使用miRNA芯片技术分析检测蜂胶黄酮PB3A处理人类结肠癌SW480细胞后miRNA表达谱的变化。通过实时荧光定量PCR方法检测miRNA-198和miRNA-296-5p的表达水平,以此来验证miRNA芯片结果的准确性和可靠性。利用miRWalk、Micro T、miRanda等12个网上数据库预测这2条miRNAs的靶基因并进行靶基因功能富集分析。结果:miRNA芯片分析结果显示,蜂胶黄酮PB3A干预24 h后结肠癌SW480细胞中差异表达倍数在2倍及以上的miRNA有267条,其中差异表达倍数达10倍及以上的miRNA有30条,28条为上调表达,2条下调表达;RT-qPCR实验结果显示miRNA-198和miRNA-296-5p的表达量趋势跟miRNA芯片结果一致,表达差异有统计学意义(P0.05)。miRNA靶基因预测发现miRNA-198有859个靶基因,miRNA-296-5p有906个靶基因;对这些可能被调控的靶基因进行Gene Class分析,结果显示miRNA-198和miRNA-296-5p的靶基因功能主要为转录因子、拷贝数变异、细胞分化、癌基因、蛋白激酶、组蛋白、转移癌基因、肿瘤抑制基因等(P0.05)。信号通路富集分析结果显示,miRNA-198靶基因显著富集于肿瘤通路、Wnt信号通路、胞吞通路、Erb B信号通路、黏着斑通路、黑素生成通路等信号通路,而miRNA-296-5p靶基因在MAPK信号通路、胞吞通路、轴突导向通路、Wnt信号通路、胰岛素信号通路、钙离子信号通路等信号通路中出现聚集(P0.05)。结论:蜂胶黄酮PB3A影响结肠癌SW480细胞的miRNA表达谱。PB3A作用下miRNA-198和miRNA-296-5p的异常表达可能参与PB3A抗结肠癌的过程。
[Abstract]:Objective: to investigate the effect of pinobanksin-3-acetatePB3A on the expression profile of microRNA-miRNAs in colon cancer SW480 cells, and to provide theoretical basis for the treatment of colon cancer and the development of targeted drugs. Methods: miRNA microarray technique was used to detect the expression profile of miRNA in human colon cancer cell line SW480 treated with propolis flavonoid PB3A. The expression levels of miRNA-198 and miRNA-296-5p were detected by real-time fluorescent quantitative PCR to verify the accuracy and reliability of miRNA chip results. The target genes of the two miRNAs were predicted by 12 online databases such as Micro TnmiRanda and the functional enrichment analysis of the two miRNAs was carried out. Results the results of microarray analysis showed that there were 267 miRNAs with a differential expression multiple of 2 times or more in colon cancer SW480 cells after 24 h intervention with propolis flavonoid PB3A. Among them, 30 miRNAs with a multiple of 10 times or more were up-regulated and 2 down-regulated expressions were detected by RT-qPCR. The results of RT-PCR showed that miRNA-198 and miRNA-296-5p showed the same tendency as those of miRNA microarray, and the results of RT-PCR showed that the expression level of miRNA-198 and miRNA-296-5p were the same as those of miRNA microarray. There were significant differences in expression of miRNA-198, 859 target genes of miRNA-296-5p and 906 target genes of miRNA-198. Gene Class analysis showed that the function of the target genes of miRNA-198 and miRNA-296-5p was mainly transcriptional factors, and the results showed that the target gene function of miRNA-198 and miRNA-296-5p was mainly transcriptional factors, and the results showed that the target genes of miRNA-198 and miRNA-296-5p were mainly transcriptional factors. Copy number variation, cell differentiation, oncogene, protein kinase, histone, metastatic oncogene, tumor suppressor gene, etc. The results of signal pathway enrichment analysis showed that the target gene of miRNA-198 was significantly enriched in the tumor pathway, such as Wnt signaling pathway, endocytosis pathway, Erb B signaling pathway, adhesion plaque pathway, melanogenesis pathway, while miRNA-296-5p target gene was located in MAPK signal pathway and cytosolic pathway. The aggregation of P0.05 was found in the signal pathways such as Wnt signaling pathway, insulin signaling pathway and calcium signal pathway. Conclusion: the expression of miRNA-198 and miRNA-296-5p in colon cancer cell line SW480 might be affected by propolis flavonoid PB3A. The abnormal expression of miRNA-198 and miRNA-296-5p may be involved in the process of PB3A anti-colon cancer.
【作者单位】：新疆大学生命科学与技术学院;
【基金】：国家自然科学基金资助项目(No.31660333)
【分类号】：R735.35

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