Wu Chang-xue, Zhang Er-yong, Yang Si-yuan, Ma Jian-yang, An Qi, Shi Ying-kang
Department of Thoracic and Cardiovascular Surgery, West China Hospital, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2008 Nov;39(6):882-5.
To investigate the expression of the transcriptive factors, GATA-4 and Nkx2.5 gene, in the course of induction of mesenchymal stem cells (MSCs) into cardiomyocytes, and analyze its molecular biological mechanism.
Rattus Mesenchymal Stem Cells were cultured in vitro and 3rd generation cells were induced in 10 micromol/L 5-azacytidine, troponin I was checked with immunohistochemistry and Western blot, GATA-4 and NkX2.5 gene were analyzed by RT-PCR in 1, 2, 3 and 4 weeks respectively.
Growth of MSCs stopped after being induced, trending to form colonies, troponin I positive expressed since 2nd week, GATA-4 and Nkx2.5 began to express in 1st week, increasing gradually, peak in 3rd week.
GATA-4 and Nkx2. 5 gene expression increased gradually in the course of stem cells differentiation, which may control the rule of cardiomyocyte transcription.
探讨转录因子GATA-4和Nkx2.5基因在间充质干细胞(MSCs)向心肌细胞诱导过程中的表达情况,并分析其分子生物学机制。
体外培养大鼠间充质干细胞,取第3代细胞用10微摩尔/升5-氮杂胞苷诱导,采用免疫组织化学和蛋白质印迹法检测肌钙蛋白I,分别于第1、2、3和4周用逆转录聚合酶链反应(RT-PCR)分析GATA-4和NkX2.5基因。
诱导后MSCs生长停止,趋于形成集落,自第2周起肌钙蛋白I呈阳性表达,GATA-4和Nkx2.5于第1周开始表达,逐渐增加,第3周达高峰。
在干细胞分化过程中,GATA-4和Nkx2.5基因表达逐渐增加,可能调控心肌细胞转录规律。