Department of Cancer Biology and Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA
Leukemia. 2012 Jul;26(7):1555-63. doi: 10.1038/leu.2012.19. Epub 2012 Jan 30.
Expression of the transcription repressor Gfi-1 is required for the maintenance of murine hematopoietic stem cells. In human cells, ectopic expression of Gfi-1 inhibits and RNA interference-mediated Gfi-1 downregulation enhances proliferation and colony formation of p210BCR/ABL expressing cells. To investigate the molecular mechanisms that may explain the effects of perturbing Gfi-1 expression in human cells, Gfi-1-regulated genes were identified by microarray analysis in K562 cells expressing the tamoxifen-regulated Gfi-1-ER protein. STAT 5B and Mcl-1, two genes important for the proliferation and survival of hematopoietic stem cells, were identified as direct and functionally relevant Gfi-1 targets in p210BCR/ABL-transformed cells because: (i) their expression and promoter activity was repressed by Gfi-1 and (ii) when constitutively expressed blocked the proliferation and colony formation inhibitory effects of Gfi-1. Consistent with these findings, genetic or pharmacological inhibition of STAT 5 and/or Mcl-1 markedly suppressed proliferation and colony formation of K562 and CD34+ chronic myelogenous leukemia (CML) cells. Together, these studies suggest that the Gfi-1STAT 5B/Mcl-1 regulatory pathway identified here can be modulated to suppress the proliferation and survival of p210BCR/ABL-transformed cells including CD34+ CML cells.
转录抑制因子 Gfi-1 的表达对于维持小鼠造血干细胞的功能是必需的。在人类细胞中,Gfi-1 的异位表达抑制了 p210BCR/ABL 表达细胞的增殖和集落形成,而 RNA 干扰介导的 Gfi-1 下调则增强了其增殖和集落形成。为了研究可能解释扰乱人类细胞中 Gfi-1 表达的分子机制,通过在表达他莫昔芬调控的 Gfi-1-ER 蛋白的 K562 细胞中进行微阵列分析,鉴定了 Gfi-1 调控的基因。STAT5B 和 Mcl-1 是造血干细胞增殖和存活的两个重要基因,被鉴定为 p210BCR/ABL 转化细胞中直接的、功能相关的 Gfi-1 靶基因,因为:(i)它们的表达和启动子活性被 Gfi-1 抑制;(ii)当它们组成性表达时,会阻断 Gfi-1 对增殖和集落形成的抑制作用。与这些发现一致,STAT5 和/或 Mcl-1 的遗传或药理学抑制显著抑制了 K562 和 CD34+慢性髓系白血病(CML)细胞的增殖和集落形成。综上所述,这些研究表明,在这里鉴定的 Gfi-1-STAT5B/Mcl-1 调控途径可以被调节,以抑制 p210BCR/ABL 转化细胞,包括 CD34+CML 细胞的增殖和存活。