Functional analysis of ETV6/ARG gene develop differentiation therapy for leukemias

ETV 6/TEL基因是ETS家族成员之一的转录因子。该基因与PDGFRB、MDS 1/EVI 1、BTL、ACS 2、STL、JAK 2、ABL、CDX 2、TRKC、AML 1、MN 1、Pax 5等多种基因融合，其中PDGFRB、ABL、JAK 2和TRKC为酪氨酸激酶（tyrosine kinase，TK）。我们认为ETV 6融合蛋白的功能分析将有助于进一步了解白血病的发病机制。我们已经鉴定了一种新的ETV 6伴侣基因ARG（ABL相关基因或ABL 2），这是从AML-M3患者中建立的细胞系中的另一种TK基因，该患者具有at（15 ; 17）（q22 ; q11.2）和at（1 ; 12）（q25 ; p13）。我们已经分析了这种酪氨酸激酶/ETV 6融合蛋白的功能，并且已经证明该蛋白赋予Ba/F3细胞IL 3非依赖性生长，以及Rat- 1细胞锚定非依赖性生长活性，表明该蛋白具有与其他酪氨酸激酶/TETV 6蛋白类似的致癌活性。进一步的研究表明，ETV 6/ARG癌蛋白 ...更多信息 通过类似的信号分子包括PI 3 K、SHC、ras-GAP和CRK-L触发与活化的ABL癌基因相关的一些相同的信号通路。ABL与多种人类恶性肿瘤的发生有关，而ARG与ABL的同源性很高，但ARG与人类恶性肿瘤的关系尚未见报道。因此，这是首次报道ARG参与人类白血病的发生，ETV 6/ARG蛋白具有致癌活性。在从M3患者建立的HT 93 Acell系中，在用全反式视黄酸和细胞因子（IL 3、GCSF、GM-CSF）培养时观察到嗜中性粒细胞、嗜酸性粒细胞和嗜碱性粒细胞的独特分化。在T-ALL细胞系中，在具有细胞因子（IL 3、G-CSF和GM-CSF）的培养物中观察到令人惊讶的向骨髓细胞的分化。因此，ETV 6/ARG蛋白可能在特定条件下与某些细胞因子一起具有分化能力。因此，我们开始研究ETV 6/ARG蛋白的分化能力。将ETV 6/ARG转染K562或HL 60细胞后，继续观察细胞形态学变化。用完全ETV 6/ARG构建体转染，未观察到细胞形态学变化。目前，我们正在用ETV 6A 5/ARG构建体进行类似的实验，该构建体具有ETV 6 ex的缺失。5.少

The ETV6/TEL gene is a transcription factor, one of the ETS family members. This gene has been reported to fuse to a variety of genes, PDGFRB, MDSl/EVI1, BTL, ACS2, STL, JAK2, ABL, CDX2, TRKC, AML1, MN1, Pax5 and so on. Among them, PDGFRB, ABL, JAK2 and TRKC are tyrosine kinases (TK). We believe that function analysis of ETV6 fusion proteins should contribute much to understanding mechanism of leukemogenesis. We already identified a novel ETV6 partner gene, ARG (ABL related gene or ABL2), another TK gene in a cell line established from an AML-M3 patient with at(15 ; 17)(q22 ; q1 1.2) and at(1 ; 12)(q25 ; p13). We have analysed function of this tyrosine kinase/ETV6 fusion protein, and have demonstrated that this protein confers IL3 independent growth to Ba/F3 cells, and anchorage independent growth activity to Rat- 1 cells, indicating that this protein possesses oncogenic activity similar to the other tyrosine kinase/TETV6 proteins. Further study has shown that the ETV6/ARG oncoprotein … More triggers some of the same signaling pathways associated with activated ABL oncogenes through similar signaling molecules including PI3K, SHC, ras-GAP and CRK-L. Although the ABL is well known to be involved in various human malignancies, ARG has not been reported to be involved in human malignancies despite of its high homology to ABL. Thus, this is the first report showing involvement of ARG in human leukemia, and that ETV6/ARG protein has oncogenic activity.ETV6/ARG fusion gene has been reported in three types of leukemias (M3, M4eo and T-ALL), so far. In the HT93Acell line established from a M3 patient, unique differentiation to neutrophils, eosinophils and basophils is observed in culture with all-trans retinoic acid and cytokines (IL3, GCSF, GM-CSF). In T-ALL cell line, a surprising differentiation to myeloid cells is observed in culture with cytokines (IL3, G-CSF, and GM-CSF). Taken together, ETV6/ARG protein is likely to possess differentiation capacity under specific condition with some cytokines. Therefore, we started to examine differentiation capacity of ETV6/ARG protein. After transfection of ETV6/ARG constructs to K562 or HL60, we continued to observe change of cell morphology. With full ETV6/ARG construct transfection, cell morphological change has not been observed. Now, we are doing the similar experiment with ETV6 A 5/ARG construct which has deletion of ETV6 ex. 5. Less