Fila-Danilow Anna, Borkowska Paulina, Paul-Samojedny Monika, Kowalczyk Malgorzata, Kowalski Jan
Department of Medical Genetics, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Sosnowiec, Poland.
Postepy Hig Med Dosw (Online). 2017 Mar 27;71(0):236-242. doi: 10.5604/01.3001.0010.3809.
Epigenetic mechanisms regulate the transcription of genes, which can affect the differentiation of MSCs. The aim of the current work is to determine how the histone deacetylase inhibitors TSA and VPA affect the expression of neuronal lineage genes in a culture of rat MSCs (rMSCs).
We analyzed the expression of early neuron marker gene (Tubb3), mature neuron markers genes (Vacht, Th, Htr2a) and the oligodendrocyte progenitor marker gene (GalC). Moreover, changes in the gene expression after three different periods of exposure to TSA and VPA were investigated for the first time.
After six days of exposition to TSA and VPA, the expression of Tubb3 and GalC decreased, while the expression of Th increased. The highest increase of VAChT expression was observed after three days of TSA and VPA treatment. A decrease in Htr2a gene expression was observed after TSA treatment and an increase was observed after VPA treatment. We also observed that TSA and VPA inhibited cell proliferation and the formation of neurospheres in the rMSCs culture.
The central findings of our study are that TSA and VPA affect the expression of neuronal lineage genes in an rMSCs culture. After exposure to TSA or VPA, the expression of early neuronal gene decreases but equally the expression of mature neuron genes increases. After TSA and VPA treatment ER of the oligodendrocyte progenitor marker decreased. TSA and VPA inhibit cell proliferation and the formation of neurospheres in rMSCs culture.
表观遗传机制调控基因转录，这会影响间充质干细胞（MSC）的分化。当前研究的目的是确定组蛋白去乙酰化酶抑制剂曲古抑菌素A（TSA）和丙戊酸（VPA）如何影响大鼠间充质干细胞（rMSC）培养物中神经谱系基因的表达。
我们分析了早期神经元标记基因（Tubb3）、成熟神经元标记基因（囊泡乙酰胆碱转运体（VAChT）、酪氨酸羟化酶（Th）、5-羟色胺受体2A（Htr2a））和少突胶质细胞祖细胞标记基因（半乳糖脑苷脂（GalC））的表达。此外，首次研究了在暴露于TSA和VPA三个不同时间段后基因表达的变化。
在暴露于TSA和VPA六天后，Tubb3和GalC的表达下降，而Th的表达增加。在TSA和VPA处理三天后观察到VAChT表达的最高增加。在TSA处理后观察到Htr2a基因表达下降，在VPA处理后观察到增加。我们还观察到TSA和VPA抑制rMSC培养物中的细胞增殖和神经球形成。
我们研究的主要发现是TSA和VPA影响rMSC培养物中神经谱系基因的表达。暴露于TSA或VPA后，早期神经元基因的表达下降，但成熟神经元基因的表达同样增加。在TSA和VPA处理后，少突胶质细胞祖细胞标记物的内质网减少。TSA和VPA抑制rMSC培养物中的细胞增殖和神经球形成。
