Overexpression of Interferon Regulatory Factor IRF1 Remodels Homeostasis of K562 Cells.
10.7534/j.issn.1009-2137.2020.01.009
- Author:
Wen-Tian WANG
1
,
2
;
Hui-Juan ZHAO
1
,
3
;
Yang YANG
1
,
2
;
Ying CHI
1
,
2
;
Lei ZHANG
1
,
4
Author Information
1. State Key Laboratory of Experimental Hematology
2. National Clinical Research Center for Blood Diseases, Tianjin Key Laboratory of Blood Disease Gene Therapy, Key Laboratory of Gene Therapy for Blood Diseases, Chinese Academy of Medical Sciences, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China.
3. National Clinical Research Center for Blood Diseases, Tianjin Key Laboratory of Blood Disease Gene Therapy, Key Laboratory of Gene Therapy for Blood Diseases, Chinese Academy of Medical Sciences, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China,Medical College, Henan University of Science and Technology, Luoyang 471023, Henan Province, China.
4. National Clinical Research Center for Blood Diseases, Tianjin Key Laboratory of Blood Disease Gene Therapy, Key Laboratory of Gene Therapy for Blood Diseases, Chinese Academy of Medical Sciences, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China,E-mail: zhanglei1@ihcams.ac.cn.
- Publication Type:Journal Article
- From:
Journal of Experimental Hematology
2020;28(1):51-55
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effects of IRF1 on the homeostasis and differentiation of K562 cells.
METHODS:Three different vectors were constructed to screen the best strategy for IRF1 overexpression. The effect of IRF1 on cell proliferation and apoptosis was explored by cell count and apoptotic surface marker detection. Likely, the effect of IRF1 on cell differentiation was analyzed by differentiational surface marker assay. Finally, the regulation mechanism at mRNA level was analyzed by RT-qPCR.
RESULTS:The single open reading frame constructed by P2A-T2A element showed the highest expression intensity, and it was the best approach to realize IRF1 enhancement. Cell counts showed that IRF1 had no significant effect on the proliferation of K562. Annexin V and 7-AAD labeling exhibited strong anti-apoptotic function of IRF1 against AraC induction. Flow cytometry revealed that IRF1 overexpression could also further increase the proportion of CD71CD235a cells. RT-qPCR confirmed its upregulation effect on CD235a and TAL1.
CONCLUSION:IRF1 enhancement alters the homeostasis characteristics of K562 cells, increases the anti-apoptotic ability and raises the potential to downstream differentiation, suggesting that IRF1 may play an important regulatory role in the hematopoietic development, including erythropoiesis.
