Nahreini Piruz, Andreatta Cynthia P, Prasad Kedar N, Toribara Neil W
Department of Gastroenterology and Hepatology, University of Colorado Health Sciences Center (UCHSC) and Denver Health Medical Center (DHMC), Denver, CO 80204, USA.
Indian J Pediatr. 2008 Oct;75(10):1009-13. doi: 10.1007/s12098-008-0176-5. Epub 2008 Sep 22.
Drug-induced differentiation is commonly used as a therapeutic modality for the treatment of neuroblastoma tumors. Increased level of cyclic adenosine 3', 5'-monophosphate (cAMP) mediates terminal differentiation in some neuroblastoma cell lines through activation of several signaling networks, including cAMP response element binding protein (CREB). Objective was to test whether cAMP-induced differentiation in a murine neuroblastoma cell line (NBP2) is partly mediated by CREB.
Fluorescent microscopy was used to document neuron-like morphological changes imparted by a constitutively active CREB (VP16CREB). Real time PCR (RT-PCR) was performed to verify changes in the expression of cAMP/CREB responsive genes.
It was found that transient expression of VP16CREB into NBP2 cells resulted in morphological changes that were characteristics of terminally differentiated neurons. Furthermore, increased expression of cAMP responsive genes was compromised in cells resisting VP16CREB-mediated differentiation.
A constitutively active CREB induces terminal differentiation in a subset of NBP2 cell population. Altered expression of cAMP responsive genes may account for differentiation resistant phenotype in NBP2 cells.
药物诱导分化通常用作治疗神经母细胞瘤肿瘤的一种治疗方式。环磷酸腺苷（cAMP）水平升高通过激活包括环磷酸腺苷反应元件结合蛋白（CREB）在内的多个信号网络，介导某些神经母细胞瘤细胞系的终末分化。目的是测试cAMP诱导的小鼠神经母细胞瘤细胞系（NBP2）分化是否部分由CREB介导。
使用荧光显微镜记录组成型活性CREB（VP16CREB）引起的神经元样形态变化。进行实时聚合酶链反应（RT-PCR）以验证cAMP/CREB反应性基因表达的变化。
发现将VP16CREB瞬时转染到NBP2细胞中会导致形态变化，这些变化是终末分化神经元的特征。此外，在抵抗VP16CREB介导的分化的细胞中，cAMP反应性基因的表达增加受到损害。
组成型活性CREB诱导NBP2细胞群体的一个亚群发生终末分化。cAMP反应性基因表达的改变可能解释了NBP2细胞中的分化抗性表型。
