论文部分内容阅读
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Taq Man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 investigate the changes of microRNA expression during the development and maturation of natural killer T cells (NKT). Methods: NKT cells were isolated from mouse thymus at different developmental stages by flow cytometry and total cellular RNA was extracted. After reverse transcription and pre-amplification, Taq Man low density microRNA expression profiling arrays were used to detect NKT development and maturation Express changes in microRNAs and use real-time PCR for further validation. Results: During the development and maturation of NKT cells, a total of 92 microRNAs changed significantly. There were 71 microRNAs with significantly increased expression, of which 36 continued to increase, while 21 with significantly reduced expression of microRNAs, of which 12 were continuously decreased. Real-time PCR of Let-7f, miR-150, miR-155, miR-223, miR-24 and miR- , The expression of miR-223 and miR-155 increased during maturation, and the trend of expression was consistent with the analysis of expression profile. CONCLUSION: The expression of NKT cells in a variety of specific microRNAs is different during development and maturation, suggesting that specific miRNAs may regulate the development and function of NKT cells.