Hatzfeld J, Li M L, Brown E L, Sookdeo H, Levesque J P, O'Toole T, Gurney C, Clark S C, Hatzfeld A
Laboratoire C.N.R.S. de Biologie Cellulaire et Moléculaire des Facteurs de Croissance, ICIG, Hôpital Paul-Brousse, Villejuif, France.
J Exp Med. 1991 Oct 1;174(4):925-9. doi: 10.1084/jem.174.4.925.
We have used antisense oligonucleotides to study the roles of transforming growth factor beta (TGF-beta) and the two antioncogenes, retinoblastoma susceptibility (Rb) and p53, in the negative regulation of proliferation of early hematopoietic cells in culture. The antisense TGF-beta sequence significantly enhanced the frequency of colony formation by multi-lineage, early erythroid, and granulomonocytic progenitors, but did not affect colony formation by late progenitors. Single cell culture and limiting dilution analysis indicated that autocrine TGF-beta is produced by a subpopulation of early progenitors. Antisense Rb but not antisense p53 yielded similar results in releasing multipotential progenitors (colony-forming unit-granulocyte/erythroid/macrophage/megakaryocyte) from quiescence. Rb antisense could partially reverse the inhibitory effect of exogenous TGF-beta. Anti-TGF-beta blocking antibodies, antisense TGF-beta, or Rb oligonucleotides all had similar effects. No additive effects were observed when these reagents were combined, suggesting a common pathway of action. Our results are consistent with the model that autocrine production of TGF-beta negatively regulates the cycling status of early hematopoietic progenitors through interaction with the Rb gene product.
我们已使用反义寡核苷酸来研究转化生长因子β（TGF-β）以及两个抗癌基因，即视网膜母细胞瘤易感基因（Rb）和p53，在体外培养的早期造血细胞增殖负调控中的作用。反义TGF-β序列显著提高了多谱系、早期红系和粒单核系祖细胞的集落形成频率，但不影响晚期祖细胞的集落形成。单细胞培养和有限稀释分析表明，自分泌TGF-β由早期祖细胞的一个亚群产生。反义Rb而非反义p53在使多能祖细胞（集落形成单位-粒细胞/红细胞/巨噬细胞/巨核细胞）从静止状态释放方面产生了类似结果。Rb反义寡核苷酸可部分逆转外源性TGF-β的抑制作用。抗TGF-β阻断抗体、反义TGF-β或Rb寡核苷酸均有类似作用。当这些试剂联合使用时未观察到相加效应，提示存在共同的作用途径。我们的结果与以下模型一致，即TGF-β的自分泌通过与Rb基因产物相互作用负调控早期造血祖细胞的循环状态。
