脂肪来源Flk-1~+间充质干细胞自我更新相关的MicroRNA

【作者】 易茹；

【导师】 章静波；

【作者基本信息】 中国协和医科大学 ， 临床医学， 2008， 博士

【摘要】 背景:随着干细胞研究的进展,研究者们报导了多个调节干细胞自我更新和增殖的基因,包括Oct-3/4、Sox2、c-Myc、Klf4和Lin28等。MicroRNA是一类非编码小RNA,通过后转录水平调控基因的表达。目前对MicroRNA在干细胞自我更新能力中的作用还知之甚少。目的:研究miR-Z在成体间充质干细胞分化早期的变化,为进一步探索其功能奠定基础。方法:诱导脂肪来源的间充质干细胞(AD-MSC)向神经、内皮、上皮、成骨、成脂方向分化,通过real-timePCR检测了诱导过程中miR-Z的表达变化,通过生物信息学方法预测了miR-Z的靶基因,并以成骨分化为例探讨了microRNA在抑制干细胞的“干性”促进其分化方面的功能。结果:miR-Z在AD-MSC各系分化的早期都上调,而其预测的靶基因SOX2在成脂、成骨、内皮分化的第3天均下调。结论:miR-Z可能通过抑制AD-MSC的干性起到促进其分化的作用。更多还原

【Abstract】 Background: With advancement of stem cell research, many regulatory factors in self-renewal and proliferation of stem cells have been reported, including Oct-3/4、Sox2、c-Myc、Klf4 and Lin28. MicroRNAs are a group of small, non-encoding RNAs, functioning through post-transcriptional regulation of target genes. Few reports have been published regarding microRNA’s function in self-renewal capacity of stem cell. Objective: The purpose of this study is to investigate the expression profile of miR-Z in early stage of adult mesenchymal stem cell. Methods: Human adipose-derived mesenchymal stem cells (AD-MSCs) were induced to neural, endothelial, epithelial, osteogenic, and adipogenic differentiation, during which expression profile of miR-Z was measured by real-time PCR. Osteogenesis was chosen as an example. Corresponding target genes of related microRNAs were predicted by bioinformatic method and their transcription and translation level in osteogenesis were confirmed by RTPCR and western blot, respectively. Results: The expression of miR-Z increased and peaked at Day 6-9 while mRNA of its predicted target SOX2 decreased at Day 3 during multi-lineage differentiation of AD-MSC. Conclusion: According to its characteristic expression profile, miR-Z may have an important function in regulating the sternness of AD-MSC.更多还原