RB and RB2/p130 genes demonstrate both specific and overlapping functions during the early steps of in vitro neural differentiation of marrow stromal stem cells
RB and RB2/p130 genes demonstrate both specific and overlapping functions during the early steps of in vitro neural differentiation of marrow stromal stem cells
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DOI：
10.1038/sj.cdd.4401499
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发表时间：
2005-01-01
影响因子：
12.4
通讯作者：
Galderisi, U
中科院分区：
文献类型：
--
作者：
Jorl, FP;Melone, MAB;Galderisi, U
骨髓基质干细胞（MSC）是干细胞样细胞，目前正在测试其在许多人类疾病的细胞治疗中的潜在用途。MSC可以分化为间充质和非间充质谱系。事实上，除了骨、软骨和脂肪之外，已经证明MSC能够分化成神经元和星形胶质细胞。RB和RB 2/p130基因参与多种系统的分化。出于这个原因，我们评估了RB和RB 2/p130在实验条件下允许MSC向神经元样表型分化的MSC的分化和凋亡中的作用。为此，我们异位表达RB或RB 2/p130，并在诱导分化为神经元样表型的大鼠原代MSC培养物中监测增殖、分化和凋亡。RB和RB 2/P130均能降低细胞增殖率。在pRb过表达细胞中，在HDAC抑制剂TSA存在下也观察到细胞生长的停滞，这表明其抗增殖活性不依赖于HDAC途径，而向pRb 2/p130过表达细胞中加入TSA可缓解生长抑制。TUNEL反应和属于Bcl-2家族的基因表达的研究表明，虽然RB保护分化的MSC免于凋亡，但与对照相比，RB 2/p130诱导凋亡增加。RB和RB 2/p130对程序性细胞死亡的影响似乎是HDAC独立的。神经分化标志物和免疫细胞化学的分子分析表明，RB 2/p130主要有助于诱导一般的神经特性和RB触发胆碱能分化。此外，RB 2/p130和RB的分化潜力似乎至少部分依赖于HDAC的活性。
Marrow stromal stem cells (MSCs) are stem-like cells that are currently being tested for their potential use in cell therapy for a number of human diseases. MSCs can differentiate into both mesenchymal and nonmesenchymal lineages. In fact, in addition to bone, cartilage and fat, it has been demonstrated that MSCs are capable of differentiating into neurons and astrocytes. RB and RB2/p130 genes are involved in the differentiation of several systems. For this reason, we evaluated the role of RB and RB2/p130 in the differentiation and apoptosis of MSCs under experimental conditions that allow for MSC differentiation toward the neuron-like phenotype. To this end, we ectopically expressed either RB or RB2/p130 and monitored proliferation, differentiation and apoptosis in rat primary MSC cultures induced to differentiate toward the neuron-like phenotype. Both RB and RB2/P130 decreased cell proliferation rate. In pRb-overexpressing cells, the arrest of cell growth was also observed in the presence of the HDAC-inhibitor TSA, suggesting that its antiproliferative activity does not rely upon the HDAC pathway, while the addition of TSA to pRb2/p130-overexpressing cells relieved growth inhibition. TUNEL reactions and studies on the expression of genes belonging to the Bcl-2 family showed that while RB protected differentiating MSCs from apoptosis, RB2/p130 induced an increase of apoptosis compared to controls. The effects of both RB and RB2/p130 on programmed cell death appeared to be HDAC-independent. Molecular analysis of neural differentiation markers and immunocytochemistry revealed that RB2/p130 contributes mainly to the induction of generic neural properties and RB triggers cholinergic differentiation. Moreover, the differentiation potentials of RB2/p130 and RB appear to rely, at least in part, on the activity of HDACs.