Analysis of Promyelocytic Leukemia in Human Embryonic Carcinoma Stem Cells During Retinoic Acid-Induced Neural Differentiation.

BACKGROUND

Promyelocytic leukemia protein () 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 gene expression in different stages of in vitro neural differentiation of NT2 cells, and to investigate the possible role of 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 mRNA and protein levels were assessed by quantitative real time RT-PCR (QRT-PCR) and Immunoblotting, respectively. Furthermore bisulfite sequencing PCR (BSP) was used to assess promoter methylation in NT2 cells and NT2 derived neuronal precursor cells (NT2.NPCs).

RESULTS

QRT-PCR results showed that, had maximum expression with significant differences in NT2 derived neuronal precursor cells relative to NT2 cells and NT2 derived neural cells (NT2.NCs). Numerous isoforms of with different intensities appeared in immunoblots of pluripotent NT2 cells, NT2.NPCs, and NT2.NCs. Furthermore, the methylation of the promoter in NT2.NCs was 2.6 percent lower than NT2 cell.

CONCLUSIONS

The observed differences in expression in different cellular stages possibly could be attributed to the fact that in each developmental state might be involved in different cell signaling machinery and different functions. The appearance of different isoforms with more intensity in neural progenitor cells; may suggest apossible role for this protein in neural development.