RUNX1-Evi-1 fusion gene inhibited differentiation and apoptosis in myelopoiesis: an in vivo study
- Shen, L., Zhu, J., Chen, F., Lin, W., Cai, J., Zhong, J., Zhong, H.
- BMC cancer 15: 970 (Journal)
- Registered Authors
- Chen, Fangyuan, Shen, Li-Jing, Zhong, Ji-Hua
- RUNX1-Evi-1, Zebrafish, Myelopoiesis, Apoptosis, Valproic acid
- MeSH Terms
- Animals, Genetically Modified
- Blotting, Western
- Cell Differentiation/genetics
- Core Binding Factor Alpha 2 Subunit/genetics*
- DNA-Binding Proteins/genetics*
- Flow Cytometry
- In Situ Hybridization
- Leukemia, Myeloid, Acute/genetics
- Oligonucleotide Array Sequence Analysis
- Oncogene Proteins, Fusion/genetics*
- Polymerase Chain Reaction
- Transcription Factors/genetics*
- 26674644 Full text @ BMC Cancer
Shen, L., Zhu, J., Chen, F., Lin, W., Cai, J., Zhong, J., Zhong, H. (2015) RUNX1-Evi-1 fusion gene inhibited differentiation and apoptosis in myelopoiesis: an in vivo study. BMC cancer. 15:970.
Background Acute myeloid leukemia (AML) 1-Evi-1 is a chimeric gene generated by the t (3; 21) (q26; q22) translocation, which leads into malignant transformation of hematopoietic stem cells by unclear mechanisms. This in vivo study aimed to establish a stable line of zebrafish expressing the human RUNX1-Evi-1 fusion gene under the control of a heat stress-inducible bidirectional promoter, and investigate its roles in hematopoiesis and hematologic malignancies.
Methods We introduced human RUNX1-Evi-1 fusion gene into embryonic zebrafish through a heat-shock promoter to establish Tg(RE:HSE:EGFP) zebrafish. Two males and one female mosaic F0 zebrafish embryos (2.1 %) were identified as stable positive germline transgenic zebrafish.
Results The population of immature myeloid cells and hematopoietic blast cells were accumulated in peripheral blood and single cell suspension from kidney of adult Tg(RE:HSE:EGFP) zebrafish. RUNX1-Evi-1 presented an intensive influence on hematopoietic regulatory factors. Consequently, primitive hematopoiesis was enhanced by upregulation of gata2 and scl, while erythropoiesis was downregulated due to the suppression of gata1. Early stage of myelopoiesis was flourishing with the high expression of pu.1, but it was inhibited along with the low expression of mpo. Microarray analysis demonstrated that RUNX1-Evi-1 not only upregulated proteasome, cell cycle, glycolysis/gluconeogenesis, tyrosine metabolism, drug metabolism, and PPAR pathway, but also suppressed transforming growth factor β, Jak-STAT, DNA replication, mismatch repair, p53 pathway, JNK signaling pathway, and nucleotide excision repair. Interestingly, histone deacetylase 4 was significantly up-regulated. Factors in cell proliferation were obviously suppressed after 3-day treatment with histone deacetylase inhibitor, valproic acid. Accordingly, higher proportion of G1 arrest and apoptosis were manifested by the propidium iodide staining.
Conclusion RUNX1-Evi-1 may promote proliferation and apoptosis resistance of primitive hematopoietic cell, and inhibit the differentiation of myeloid cells with the synergy of different pathways and factors. VPA may be a promising choice in the molecular targeting therapy of RUNX1-Evi-1-related leukemia.
Genes / Markers
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Engineered Foreign Genes