科研之友
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摘要
Background: Promyelocytic leukemia protein (PML) is a tumor suppressor protein that is involved in myeloid cell differentiation in response to retinoic acid (RA). In addition, RA acts as a natural morphogen in neural development. Objectives: This study aimed to examine PAIL gene expression in different stages of in vitro neural differentiation of NT2 cells, and to investigate the possible role of PAIL in pluripotency and/or neural development. Materials and Methods: RA was used as a neural inducer for in vitro neural differentiation of NT2 cells, During this process PAIL mRNA and protein levels were assessed by quantitative real time RT-PCR (QRT-PCR) and Immunoblotting, respectively. Furthermore biosulfite sequencing PCR (BSP) was used to assess PAIL promoter methylation in NT2 cells and NT2 derived neuronal precursor cells (NT2.NPCs). Results: QRT-PCR results showed that. PAIL had maximum expression with significant differences in NT2 derived neuronal precursor cells relative to NT2 cells and NT2 derived neural cells (NTINCs). Numerous isoforms of PML with different intensities appeared in immunoblots of pluripotent NT2 cells, NT2.NPCs. and NT2,NCs. Furthermore, the methylation of the PML promoter in NT2.NCs was 2.6 percent lower than NT2 cell. Conclusions: The observed differences in PML expression in different cellular stages possibly could be attributed to the fact that PAIL in each developmental state might be involved in different cell signaling machinery and different functions. The appearance of different PML isoforms with more intensity in neural progenitor cells; may suggest apossible role for this protein in neural development.
- 出版日期2016
