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| Status |
Public on Mar 29, 2016 |
| Title |
Analysis of the effects of ZNF16 on gene expression in k562 cells. |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
We previously characterized zinc finger protein gene HZF1 (ZNF16) and demonstrated its important roles in erythroid and megakaryocytic differentiation of K562 cells by loss-function assay. However its effect in erythroid and megakaryocytic differentiation of hematopoietic stem/progenitor cells (HSPCs) and the mechanisms by which it functions have not been understood. In this study, we detected up-regulation of ZNF16 during erythroid and megakaryocytic differentiation of K562 cells and normal CD34+ HSPCs, and demonstrated that ZNF16 promotes erythroid and megakaryocytic differentiation by gain-of-function and loss-of-function experiments. Gene expression profiling by mRNA array and PCR validation in the K562 transforments with ZNF16 over-expression suggested that cell division cycle-associated 7-like gene (JPO2) and v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog gene (c-KIT) were among the genes regulated possibly by ZNF16. Luciferase reporter assay and Chromatin Immunoprecipitation demonstrated that ZNF16 binds to JPO2 and c-KIT promoters and inhibits their expression in K562 cells. A significant decrease of JPO2 and c-KIT levels was observed during erythroid and megakaryocytic differentiation of K562 and CD34+ cells. The knockdown of either JPO2 or c-KIT partially rescued the differentiation inhibition caused by ZNF16 knockdown. We also found that ZNF16 inhibits c-KIT/c-Raf/MEK/ERK/c-Jun/HEY1 signal pathways, which finally up-regulated expression of GATA1, a central regulator of erthroid and megakaryocyte differentiation. By lentivirus-mediated gene transfer, we demonstrated that enforced expression and knockdown of ZNF16 in HSPCs down-regulated and up-regulated expression of its targets respectively. Our data collectively demonstrate that ZNF16 promotes erythropoiesis and megakaryocytopoiesis via its regulation on JPO2 and c-KIT.
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| Overall design |
4 samples are analyzed, H4-1 and H4-2 are two stable K562 transductants with ZNF16 over-expression, PC1 and PC2 are stable control K562 transductants. Stable K562 transductants were obtained for RNA extraction and hybridization on an Illumina HumanHT-12 V4.0 expression beadchipe xpression Array platform.
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| Contributor(s) |
Chen J, Zhang J |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| |
| Submission date |
May 02, 2013 |
| Last update date |
Mar 29, 2016 |
| Contact name |
Junwu Zhang |
| E-mail(s) |
zhangjunwupumc@sina.cn
|
| Organization name |
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences,
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| Street address |
5 Dong Dan San Tiao
|
| City |
Bei Jing |
| ZIP/Postal code |
100005 |
| Country |
China |
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| Platforms (1) |
| GPL10904 |
Illumina HumanHT-12 V4.0 expression beadchip (gene symbol) |
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| Samples (4)
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| GSM1132996 |
Stable K562 transductants _ ZNF16 over-expression_rep1 |
| GSM1132997 |
Stable K562 transductants _ ZNF16 over-expression_rep2 |
| GSM1132998 |
Stable K562 transductants_ control_ rep1 |
| GSM1132999 |
Stable K562 transductants_ control_ rep2 |
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| Relations |
| BioProject |
PRJNA201064 |
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