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  

Expression of miRNA-198 and miRNA-296-5P in PB3A-treated colon cancer cell line and its function prediction

作　　者：胡尼其古丽.阿巴克 玛依努尔.达吾提 依米提.热合曼 阿依努尔.玉苏普 吾热娅提古丽.克维尔 买日江古丽.阿布力提甫 

机构地区：[1]新疆大学生命科学与技术学院,新疆乌鲁木齐830046

出　　处：《中国病理生理杂志》2017年第7期1317-1322,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.31660333)

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(P<0.05)。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(P<0.05)。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(P<0.05)。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。AIM： To explore the effect of pinobanksin-3-acetate （PB3A） on microRNA （miRNA） expression profile of human colon cancer cells for providing new methods of treatment of colon cancer and development of targeted drug. METHODS ： The method of miRNA expression profiling was used to observe the miRNA differential expression in human colon cancer SW480 cells after treated with PB3A. The expression of miRNA-198 and miRNA-296-5p in the SW480 cells was detected by RT-qPCR. The network databases of miRWalk, MicroT, miRanda and so on were used to predict the target genes regulated by these miRNAs, and pathway significant enrichment analysis was performed. RESULTS ： miRNA microarray analysis showed that after treated with propolis flavonoid PB3A for 24 h, 267 miRNAs with differential expres-sion twice or more in the SW480 cells were observed. Among them, there were 30 miRNAs with 10-fold or more differential expression, in which 28 were up-regulated and 2 were down-regulated. The results of RT-qPCR showed that the expression levels of miRNA-198 and miRNA-296-5p were consistent with the results of miRNA microarray analysis, and the difference was statistically significant （ P 〈 0. 05 ） . Bioinformatic analysis revealed that miRNA-198 has 859 target genes, and miRNA-296-5p has 906 target genes. The target genes of miRNA-198 were clustered in pathways in cancer, axon guidance,Wnt signaling pathway, regulation of actin cytoskeleton, insulin signaling pathway and MAPK signaling pathway, while the target genes of miRNA-296-5p were clustered in axon guidance, Wnt signaling pathway, MAPK signaling path-way ,endocytosis, melanogenesis, insulin signaling pathway and calcium signaling pathway. CONCLUSION ： Propolis flavonoid PB3A affects the expression of miRNA in colon cancer SW480 cells. The abnormal expression of miRNA-198 and miRNA-296-5p may be involved in the inhibitory effect of PB3A on colon cancer.

关 键 词：短叶松素-3-乙酸脂 结肠癌SW480细胞 微小RNA-198 微小RNA-296-5p 

分 类 号：R730.23[医药卫生—肿瘤]