Department of Histology and Embryology, Harbin Medical University, Harbin, China.
Mol Biol Rep. 2010 Mar;37(3):1197-202. doi: 10.1007/s11033-009-9489-3. Epub 2009 Mar 5.
Recently, differentiated somatic cells had been reprogrammed to pluripotential state in vitro, and various tissue cells had been elicited from those cells. Epigenetic modifications allow differentiated cells to perpetuate the molecular memory needed for the cells to retain their identity. DNA methylation and histone deacetylation are important patterns involved in epigenetic modification, which take critical roles in regulating DNA expression. In this study, we dedifferentiated NIH/3T3 fibroblasts by 5-aza-2-deoxycytidine (5-aza-dC) and Trichstatin A (TSA) combination, and detected gene expression pattern, DNA methylation level, and differentiation potential of reprogrammed cells. As the results, embryonic marker Sox2, klf4, c-Myc and Oct4 were expressed in reprogrammed NIH/3T3 fibroblasts. Total DNA methylation level was significant decreased after the treatment. Moreover, exposure of the reprogrammed cells to all trans-retinoic acid (RA) medium elicited the generation of neuronal class IIIbeta-tubulin-positive, neuron-specific enolase (NSE)-positive, nestin-positive, and neurofilament light chain (NF-L)-positive neural-like cells.
最近，已在体外将分化的体细胞重编程为多能状态，并且已经从这些细胞中诱发出各种组织细胞。表观遗传修饰允许分化细胞保持其身份所需的分子记忆。DNA 甲基化和组蛋白去乙酰化是涉及表观遗传修饰的重要模式，它们在调节 DNA 表达中起着关键作用。在这项研究中，我们通过 5-氮杂-2′-脱氧胞苷（5-aza-dC）和 Trichstatin A（TSA）组合使 NIH/3T3 成纤维细胞去分化，并检测了重编程细胞的基因表达模式、DNA 甲基化水平和分化潜能。结果表明，胚胎标记物 Sox2、klf4、c-Myc 和 Oct4 在重编程的 NIH/3T3 成纤维细胞中表达。治疗后总 DNA 甲基化水平显著降低。此外，将重编程细胞暴露于全反式视黄酸（RA）培养基中可诱导神经元类 IIIβ-微管蛋白阳性、神经元特异性烯醇化酶（NSE）阳性、巢蛋白阳性和神经丝轻链（NF-L）阳性的神经样细胞生成。
