目的应用miRNAs芯片筛选中波紫外线(UVB)照射的NIH3T3细胞表达miRNAs,并寻找差异表达miRNAs调控的靶基因。方法 miRNAs微阵列技术筛选UVB照射的NIH3T3细胞差异表达miRNAs,同时运用miRNAs特异的引物组对差异表达的miRNAs进行荧光定量RT-PCR验证。结果 NIH3T3细胞受不同剂量UVB照射后,MTT结果显示细胞生存率明显下降。miRNAs芯片结果显示,实验组细胞与对照组细胞差异表达的miRNA共30个(占11%),其中27个表达上调,3个表达下调。mmu-miR-365和mmu-miR-21显著上调,而mmu-miR-465在不同的时间点都下调。其差异表达均在2倍以上;mmu-miR-let-7a、mmu-miR-24、mmu-miR-376b和mmu-miR-21的实时荧光定量RT-PCR验证结果与芯片表达结果基本一致。结论 miRNA在UVB照射的NIH3T3细胞中有差异表达,提示miRNA可能参与UVB照射后NIH3T3细胞内的多种信号调控途径。
Objective To investigate the differentially expressed miRNAs and their target genes in NIH3T3 cells after UVB irradiation,that is useful for further studies on functions of miRNAs in UVB induced Signal transduction pathway. Methods MicroRNA microarray was used to investigate the differentially expressed miRNAs in NIH3T3 cells after UVB irradiation,and the discovered miRNAs were confirmed by real time RT-PCR assay. Results MicroRNA microarray showed that 30 miRNAs were differentially expressed,accounting for 11% ,in which 27 up-regulated,3 down-regulated. Interestingly,the express level of mmu-miR-365 and mmu-miR-21 were more than six,while mmu-miR-465 showed low levels of expression at all time points. The expression levels of Mmu-miR-let-7a,mmu-miR-24,mmumiR-376b and mmu-miR-21 were accordant with the results from real time RT-PCR. Conclusion There is a differential expression of miRNAs in NIH3T3 after UVB irradiation,which suggests that miRNAs might involve in several cell signal pathways of NIH3T3 cells after exposure to UVB.