Condorelli G L, Tocci A, Botta R, Facchiano F, Testa U, Vitelli L, Valtieri M, Croce C M, Peschle C
Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107-5541, USA.
Mol Cell Biol. 1997 May;17(5):2954-69. doi: 10.1128/MCB.17.5.2954.
The TAL-1 gene specifies a basic helix-loop-helix domain (bHLH) transcription factor, which heterodimerizes with E2A gene family proteins. tal-1 protein is abnormally expressed in the majority of T-cell acute lymphoblastic leukemias (T-ALLs). tal-1 is expressed and plays a significant role in normal erythropoietic differentiation and maturation, while its expression in early myeloid differentiation is abruptly shut off at the level of late progenitors/early differentiated precursors (G. L. Condorelli, L. Vitelli, M. Valtieri, I. Marta, E. Montesoro, V. Lulli, R. Baer, and C. Peschle, Blood 86:164-175, 1995). We show that in late myeloid progenitors (the phenotypically normal murine 32D cell line) and early leukemic precursors (the human HL-60 promyelocytic leukemia cell line) ectopic tal-1 expression induces (i) a proliferative effect under suboptimal culture conditions (i.e., low growth factor and serum concentrations respectively), via an antiapoptotic effect in 32D cells or increased DNA synthesis in HL-60 cells, and (ii) a total or marked inhibitory effect on differentiation, respectively, on granulocyte colony-stimulating factor-induced granulopoiesis in 32D cells or retinoic acid- and vitamin D3-induced granulo- and monocytopoiesis in HL-60 cells. Furthermore, experiments with 32D temperature-sensitive p53 cells indicate that aberrant tal-1 expression at the permissive temperature does not exert a proliferative effect but causes p53-mediated apoptosis, i.e., the tal-1 proliferative effect depends on the integrity of the cell cycle checkpoints of the host cell, as observed for c-myc and other oncogenes. tal-1 mutant experiments indicate that ectopic tal-1 effects are mediated by both the DNA-binding and the heterodimerization domains, while the N-terminally truncated tal-1 variant (M3) expressed in T-ALL malignant cells mimics the effects of the wild-type protein. Altogether, our results (i) indicate proliferative and antidifferentiative effects of ectopic tal-1 expression, (ii) shed light on the underlying mechanisms (i.e., requirement for the integrity of the tal-1 bHLH domain and cell cycle checkpoints in the host cell, particularly p53), and (iii) provide new experimental models to further investigate these mechanisms.
TAL-1基因编码一种碱性螺旋-环-螺旋结构域(bHLH)转录因子,它与E2A基因家族蛋白形成异二聚体。tal-1蛋白在大多数T细胞急性淋巴细胞白血病(T-ALL)中异常表达。tal-1在正常红细胞生成分化和成熟过程中表达并发挥重要作用,而其在早期髓系分化中的表达在晚期祖细胞/早期分化前体细胞水平突然关闭(G. L. 孔多雷利、L. 维泰利、M. 瓦尔蒂耶里、I. 马尔塔、E. 蒙特索罗、V. 卢利、R. 贝尔和C. 佩施勒,《血液》86:164 - 175,1995)。我们发现,在晚期髓系祖细胞(表型正常的小鼠32D细胞系)和早期白血病前体细胞(人HL-60早幼粒细胞白血病细胞系)中,异位tal-1表达诱导:(i)在次优培养条件下(即分别为低生长因子和血清浓度)产生增殖效应,在32D细胞中通过抗凋亡作用,或在HL-60细胞中通过增加DNA合成;(ii)分别对分化产生完全或显著的抑制作用,在32D细胞中对粒细胞集落刺激因子诱导的粒细胞生成,或在HL-60细胞中对视黄酸和维生素D3诱导的粒细胞和单核细胞生成。此外,对32D温度敏感p53细胞的实验表明,在允许温度下异常的tal-1表达不产生增殖效应,但会导致p53介导的凋亡,即tal-1的增殖效应取决于宿主细胞周期检查点的完整性,这与c-myc和其他癌基因的情况相同。tal-1突变实验表明,异位tal-1效应由DNA结合结构域和异二聚化结构域介导,而在T-ALL恶性细胞中表达的N端截短的tal-1变体(M3)模拟野生型蛋白的效应。总之,我们的结果:(i)表明异位tal-1表达的增殖和抗分化效应;(ii)揭示了潜在机制(即tal-1 bHLH结构域和宿主细胞周期检查点完整性的要求,特别是p53);(iii)提供了新的实验模型以进一步研究这些机制。