人脐带间充质干细胞向成骨细胞及脂肪细胞分化过程中nm23-H1基因的表达

Expression of nm23-H1 during differentiation of human umbilical cord mesenchymal stem cells into adipocytes and osteoblasts in vitro

背景:nm23-h1基因已被证实与多种细胞的增殖、分化、转移密切相关。目的:观察人脐带间充质干细胞在体外向成脂及成骨方向诱导分化的过程中nm23-H1基因的表达变化。方法:采用组织块贴壁法从脐带中分离培养人间充质干细胞,流式细胞术鉴定其表型,之后分别加入成脂肪诱导剂和成骨诱导剂进行体外诱导分化。对照组采用不含诱导因子的培养液进行培养。结果与结论:在成脂诱导过程的第4天,nm23-H1 mRNA的表达上调,至第7天达到最高(P<0.01),其后开始逐渐恢复,直至第14天与对照组相比无显著差异(P>0.05)。而在成骨诱导过程中,nm23-H1 mRNA则一直为低表达状态(P<0.01),直至第28天恢复为对照组水平(P>0.05)。结果显示nm23-H1基因表达在成脂分化前期发生上调,而在成骨分化过程中一直下调。

BACKGROUND： It has been proved that nm23-H1 gene has a great infulence on proliferation,differentiation and transfering of many kinds of cells.OBJECTIVE： To study on the expression of nm23-H1 gene during differentiation of human umbilical cord mesenchymal stem cells （hUCMSCs） into adipocytes and osteoblasts in vitro.METHODS： The hUCMSCs were isolated from human umbilical cord by tissue adherence,and a flow cytometry was used for immunophenotype identification.Then the 3rd passage cells were induced to differentiate into adipocytes and osteoblasts,characterized by oil red O staining and Alizarin red staining respectively.The expression of nm23-H1 mRNA was detected by real time quantitative RT-PCR during the adipogenic and osteogenic differentiation.RESULTS AND CONCLUSION： During the adipogenic differentiation,the nm23-H1 expression was increased in mRNA level after the induction on 4 days,and reached to the highest level on 7 days （P 0.01）,and then was gradually decreased on 14 days to the primary level which had no significant changes compared with untreated control （P 0.05）.During the osteogenic differentiation,the nm23-H1 expression in mRNA level was downregulated （P 0.01） until maturation on 28 days （P 0.05）.nm23-H1 gene has shown a regulatory changes during the differerntiation of hUCMSCs into adipocytes and osteoblasts,this might imply that nm23-H1 gene make a contribution to the osteogenic and adipogenic differentiation.