All‐trans‐retinoid acid induces the differentiation of P19 cells into neurons involved in the PI3K/Akt/GSK3β signaling pathway

Abstract

The pluripotent mouse embryonal carcinoma cell line P19 is widely used as a model for research on all‐trans‐retinoid acid (RA)‐induced neuronal differentiation; however, the signaling pathways involved in this process remain unclear. This study aimed to reveal the molecular mechanism underlying the RA‐induced neuronal differentiation of P19 cells. Real‐time quantitative polymerase chain reaction and Western blot analysis were used to determine the expression of neuronal‐specific markers, whereas flow cytometry was used to analyze cell cycle and cell apoptosis. The expression profiles of messenger RNAs (mRNAs) in RA‐induced neuronal differentiation of P19 cells were analyzed using high‐throughput sequencing, and the functions of differentially expressed mRNAs (DEMs) were determined by bioinformatics analysis. RA induced an increase in both class III β‐tubulin (TUBB3) and neurofilament medium (NEFM) mRNA expression, indicating that RA successfully induces neuronal differentiation of P19 cells. Cell apoptosis was not affected; however, cell proliferation decreased. We found 4117 DEMs, which were enriched in the phosphoinositide 3‐kinase/protein kinase B (PI3K/Akt) signaling pathway, Wnt signaling pathway, and cell cycle. Particularly, a few DEMs could be identified in the PI3K/Akt signaling pathway networks, such as PI3K, Akt, glycogen synthase kinase‐3β (GSK3β), cyclin‐dependent kinase 4 (CDK4), P21, and Bax. RA significantly increased the protein expression of PI3K, Akt, phosphorylated Akt, GSK3β, phosphorylated GSK3β, CDK4, and P21, but it reduced Bax protein expression. The Akt inhibitor affected the increase of TUBB3 and NEFM mRNA expression in RA‐induced P19 cells. The molecular mechanism underlying the RA‐induced neuronal differentiation of P19 cells is potentially involved in the PI3K/Akt/GSK3β signaling pathway. The decreased cell proliferation ability of neuronally differentiated P19 cells could be associated with the expression of cell cycle proteins.

Cannabidiol Promotes Neuronal Differentiation Using Akt and Erk Pathways Triggered by Cb1 Signaling

“…After performing the Western blot, shown in Figure 5 f, the phosphorylation of AKT was present only at 10 µM, suggesting the effect was mediated by CB1 at 10 µM. AKT is involved in many vital cell processes, such as proliferation and differentiation, and studies correlate the presence of p-AKT as necessary to achieve differentiation, also in retinoic-acid induced differentiation [ 23 , 32 ].…”

Circular RNA circ‐RCCD promotes cardiomyocyte differentiation in mouse embryo development via recruiting YY1 to the promoter of MyD88

“…At present, three pluripotent cell lines have been isolated from living organisms, namely embryonic tumour cells, embryonic stem cells and embryonic germ cell lines 25,26 . P19 cells, derived from C3H/HE malformed neoplasms, are one of the most versatile embryonic neoplasms 27 . Cardiomyocytes induced by differentiation of P19 cells exhibit the physiological and biochemical characteristics of early embryonic myocardium 28 .…”

“…25 , 26 P19 cells, derived from C3H/HE malformed neoplasms, are one of the most versatile embryonic neoplasms. 27 Cardiomyocytes induced by differentiation of P19 cells exhibit the physiological and biochemical characteristics of early embryonic myocardium. 28 Compared with the model established by other stem cells for inducing differentiation of cardiomyocytes, P19 cells model has the best intercellular homogeneity, which is the most beneficial for analysing the signal regulation mechanism in the process of cardiomyocyte differentiation.…”

HPLC-MS/MS Shows That the Cellular Uptake of All-Trans-Retinoic Acid under Hypoxia Is Downregulated by the Novel Active Agent 5-Methoxyleoligin

“…Vitamin A and its derivatives play major roles in numerous physiological processes, including signalling pathways; regulation of the energy metabolism; cellular proliferation and differentiation; and immune system function, growth, and development, while also maintaining health and eyesight [ 1 , 2 , 3 , 4 , 5 , 6 ]. Dietary uptake of vitamin A in the form of retinol or retinyl esters results in a cascade of enzymatically catalyzed modifications to retinol that yield its corresponding aldehyde and eventually retinoic acid.…”