Blood Cancer J. 2012 May;2(5):e71. doi: 10.1038/bcj.2012.16. Epub 2012 May 11.
The c-Myb gene encodes the p75(c-Myb) isoform and less-abundant proteins generated by alternatively spliced transcripts. Among these, the best known is p(c-Mybex9b), which contains 121 additional amino acids between exon 9 and 10, in a domain involved in protein-protein interactions and negative regulation. In hematopoietic cells, expression of p(c-Mybex9b) accounts for 10-15% of total c-Myb; these levels may be biologically relevant because modest changes in c-Myb expression affects proliferation and survival of leukemic cells and lineage choice and frequency of normal hematopoietic progenitors. In this study, we assessed biochemical activities of p(c-Mybex9b) and the consequences of perturbing its expression in K562 and primary chronic myeloid leukemia (CML) progenitor cells. Compared with p75(c-Myb), p(c-Mybex9b) is more stable and more effective in transactivating Myb-regulated promoters. Ectopic expression of p(c-Mybex9b) enhanced proliferation and colony formation and reduced imatinib (IM) sensitivity of K562 cells; conversely, specific downregulation of p(c-Mybex9b) reduced proliferation and colony formation, enhanced IM sensitivity of K562 cells and markedly suppressed colony formation of CML CD34(+) cells, without affecting the levels of p75(c-Myb). Together, these studies indicate that expression of the low-abundance p(c-Mybex9b) isoform has an important role for the overall biological effects of c-Myb in BCR/ABL-transformed cells.
c-Myb 基因编码 p75(c-Myb)同工型和通过可变剪接转录本产生的较少丰度的蛋白质。其中,最著名的是 p(c-Mybex9b),它在涉及蛋白质-蛋白质相互作用和负调控的区域包含外显子 9 和 10 之间的 121 个额外氨基酸。在造血细胞中,p(c-Mybex9b)的表达占 c-Myb 的 10-15%;这些水平可能具有生物学相关性,因为 c-Myb 表达的适度变化会影响白血病细胞的增殖和存活以及正常造血祖细胞的谱系选择和频率。在这项研究中,我们评估了 p(c-Mybex9b)的生化活性以及在 K562 和原发性慢性髓系白血病 (CML)祖细胞中干扰其表达的后果。与 p75(c-Myb)相比,p(c-Mybex9b)在反式激活 Myb 调节的启动子方面更稳定且更有效。p(c-Mybex9b)的异位表达增强了 K562 细胞的增殖和集落形成,并降低了伊马替尼 (IM) 的敏感性;相反,p(c-Mybex9b)的特异性下调降低了 K562 细胞的增殖和集落形成,增强了 K562 细胞对 IM 的敏感性,并显著抑制了 CML CD34(+)细胞的集落形成,而不影响 p75(c-Myb)的水平。总之,这些研究表明低丰度 p(c-Mybex9b)同工型的表达对于 c-Myb 在 BCR/ABL 转化细胞中的整体生物学效应具有重要作用。