Guo X Y, Cuillerot J M, Wang T, Wu Y, Arlinghaus R, Claxton D, Bachier C, Greenberger J, Colombowala I, Deisseroth A B
Department of Internal Medicine and the Gene Therapy Program of the Yale Cancer Center, Yale University School of Medicine, New Haven, Connecticut 06520-8032, USA.
Oncogene. 1998 Aug 20;17(7):825-33. doi: 10.1038/sj.onc.1201999.
We first showed that the introduction of a bcr-abl transcription unit into the 32D murine myeloid cell line (P210bcrabl32D) converts this cell line from an IL3 dependent cell line to an IL3 growth independent cell line. We next cloned a fragment of the bcr-abl cDNA, which codes for the bcr oligomerization domain and neighboring regions. To test for a transformation inhibitory effect of this oligomerization inhibitory peptide transcription unit on the p210bcr-abl mediated IL3 independent growth of the P210bcrabl32D cell line, we transiently co-electroporated into the growth factor dependent 32D cells, mixtures of plasmids which contained varying ratios of the plasmid expression vectors for the bcr oligomerization inhibitory peptide along with a smaller amount of the plasmid expression vector for the full length p210bcr-abl. (The P210bcr-abl protein converts the 32D from a growth factor dependent into a growth factor independent cell line.) We then showed that the oligomerization domain containing fragment from the bcr and bcr-abl proteins, can be used to inhibit the IL3 independent growth of p210bcr-abl positive 32D cells. These studies may be of eventual interest for those investigators whose goal is to design molecular therapeutic approaches to CML based on the use of peptidomimetic chemical functionalities, which mimic the structure and the inhibitory binding properties of the oligomerization domain containing fragment so as to inhibit the transforming function of the P210bcr-abl oncoprotein.
我们首先证明，将bcr-abl转录单位引入32D小鼠髓系细胞系（P210bcrabl32D）可使该细胞系从依赖白细胞介素3（IL3）的细胞系转变为不依赖IL3生长的细胞系。接下来，我们克隆了一段bcr-abl cDNA片段，该片段编码bcr寡聚化结构域及相邻区域。为了测试这种寡聚化抑制肽转录单位对P210bcrabl32D细胞系中p210bcr-abl介导的不依赖IL3生长的转化抑制作用，我们将含有不同比例bcr寡聚化抑制肽质粒表达载体以及少量全长p210bcr-abl质粒表达载体的质粒混合物，瞬时共电穿孔导入依赖生长因子的32D细胞中。（P210bcr-abl蛋白可使32D细胞从依赖生长因子转变为不依赖生长因子的细胞系。）然后我们证明，来自bcr和bcr-abl蛋白的含寡聚化结构域片段可用于抑制p210bcr-abl阳性32D细胞的不依赖IL3生长。对于那些旨在基于使用拟肽化学官能团设计慢性粒细胞白血病分子治疗方法的研究人员来说，这些研究最终可能会引起他们的兴趣，这些拟肽化学官能团模仿含寡聚化结构域片段的结构和抑制性结合特性，从而抑制P210bcr-abl癌蛋白的转化功能。
