NKT细胞发育过程中microRNA表达谱变化

摘要

目的:探讨自然杀伤性T细胞(NKT)发育、成熟过程中microRNA表达谱变化.方法:采用流式细胞仪分选小鼠胸腺不同发育阶段NKT细胞,提取细胞总RNA,经反转录和预扩增后利用TaqMan低密度microRNA表达谱分析阵列检测NKT发育、成熟过程中发生表达变化的microRNAs,并采用real-time PCR进一步验证.结果:NKT细胞发育、成熟过程中,共有92个microRNAs表达发生显著变化.表达显著增加的microRNAs有71个,其中有36个表达持续增加;而表达显著降低的microRNAs有21个,其中有12个表达持续降低.选取Let-7f、miR-150、miR-155、miR-223、miR-24和miR-29进行real-time PCR,发现Let-7f、miR-150、miR-24、miR-29在NKT细胞发育、成熟过程中表达增加,而miR-223和miR-155表达降低,其表达变化趋势与表达谱分析一致.结论:NKT细胞发育、成熟过程中伴随大量特异microRNAs的不同表达变化,提示特异microRNA调控NKT细胞的发育和功能.%Objective:To explore the microRNA profile changes in the development of NKT cells.Methods: Differently developmental stage of NKT cells in mouse thymus were sorted by flow cytometry.Total RNA were extracted,reversely transcribed and pre-amplified.TaqMan low density microRNA assay and single real-time PCR were applied to detect the expression changes of microRNAs in the developmental process of NKT cells.Results: There were total 92 microRNAs whose expression changed significantly during the development and maturation of NKT cells.Among them,increasly expressed microRNAs were 71,including 36 microRNAs whose expression continuously increased;decreasly expressed microRNAs were 21,including 12 microRNAs whose expression continuously decreased.In addition,single real-time PCR analysis showed that the expression of Let-7f,miR-150,miR-24,miR-29 increased,while the expression of miR-223 and miR-155 decreased during the development and maturation of NKT cells.Conclusion: NKT development and maturation is accompanied by expression changes of large amount of microRNAs,indicating that specific microRNA regulates NKT development and function.