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目的研究细胞编码miRNA在轮状病毒复制中的作用,并分析轮状病毒感染对于细胞编码miRNA种类及表达的影响,结合预测初步寻找具有重要功能的单个miRNA。方法首先,利用RNAi技术成功干扰MA104细胞中miRNA生成必须的Dicer酶,利用FFA法比较Dicer干扰组与阴性对照组感染后子病毒的滴度。然后,利用miRNA微芯片技术分析MA104细胞在2株轮状病毒SA11与Wa株感染后miRNA表达变化,通过聚类分析研究变化规律。结果干扰组MA104细胞的子病毒滴度显著增高。MA104细胞编码miRNA在2株轮状病毒感染后,约有200个miRNA有不同程度的变化。结合计算机预测,初步锁定miR-512-5p,miR-490,let-7c等10个miRNA可能在轮状病毒复制中具有功能。结论轮状病毒调节其感染细胞编码的miRNA的表达谱,反过来,细胞的Dicer及细胞编码miRNA影响轮状病毒复制。
Objective To investigate the role of cell-encoded miRNAs in rotavirus replication and to analyze the effect of rotavirus infection on the types and expression of miRNAs encoding for cells, and to search for a single miRNA with important functions. Methods Firstly, the RNAi technique was used to successfully interfere with the Dicer enzyme in the miRNA production in MA104 cells. The titer of the virus in the Dicer interference group and the negative control group was compared by FFA. Then miRNA microarray was used to analyze the miRNA expression of MA104 cells infected by two rotavirus strains SA11 and Wa. The clustering analysis was used to study the changes of miRNA expression. Results The sub-virus titers of MA104 cells in the interference group were significantly increased. MA104 cells encode miRNA There are about 200 miRNAs varying degrees of change after 2 rotavirus infections. Combined with computer prediction, 10 miRNAs such as miR-512-5p, miR-490 and let-7c may be locked in rotavirus replication. Conclusion Rotavirus regulates the expression profile of miRNAs encoded by infected cells, and in turn, the cellular Dicer and cell-encoded miRNAs affect rotavirus replication.