摘要
本研究旨在探讨小鼠骨髓间充质干细胞(mesenchymal stem cells,MSC)中转录因子RUNX1对WNT5A转录活性的调控,探索骨髓造血微环境对造血干细胞调控的机制。体外分离、培养、鉴定小鼠骨髓MSC;利用RT-PCR方法检测小鼠骨髓MSC中RUNX1的表达;利用染色质免疫共沉淀研究RUNX1与WNT5A启动子之间的直接相互作用;利用逆转录病毒表达系统将RUNX1导入小鼠间充质干细胞中,用RT-PCR的方法检测基因的表达,研究RUNX1对WNT5A的转录调节作用。结果发现:体外分离培养得到的MSC经脂肪、成骨、软骨诱导后,分别用油红O、von Kossa、甲苯胺蓝染色呈阳性。RUNX1表达于小鼠骨髓来源MSC中,RUNX1能与WNT5A启动子直接结合,并对WNT5A具有转录激活作用。结论:小鼠骨髓MSC中表达RUNX1,WNT5A是RUNX1的一个下游靶基因,其转录活性受RUNX1调控。
This study was purposed to investigate the effect of RUNX1 on transcription activity of WNT5A promoter in mouse bone marrow derived mesenchymal stem cells（MSC）,and to explore the mechanism by which bone marrow environments regulate MSC.RT-PCR was used to detect the expression of RUNX1 in MSC isolated from mouse bone marrow and cultured in vitro;the chromatin immunoprecipitation（ChIP） was used to investigate the direct in vivo interaction between the RUNX1 and WNT5A promoter;retrovirus system was utilized to introduce the RUNX1 gene into MSC to detect the regulation of RUNX1 on the transcription activity of WNT5A promoter.The results showed that mouse bone marrow derived MSC was positive for Oil Red O,van Kossa and toluidine blue staining respectively and RUNX1 expressed in MSC.WNT5A promoter could be bound by RUNX1,and the expression level of WNT5A was enhanced with the increase of RUNX1.It is concluded that RUNX1 expresses in mouse bone marrow derived MSC,WNT5A is a direct target gene of RUNX1 and its transcriptional activity is regulated by RUNX1.
出处
《中国实验血液学杂志》
CAS
CSCD
2011年第5期1200-1203,共4页
Journal of Experimental Hematology
基金
国家自然科学基金资助项目(编号30800426
30730043
81070393)
