Effects of NRG4 on Proliferation, Apoptosis and Cell Cycle of Acute Myeloid Leukemia Cells

doi:10.19586/j.2095-2341.2022.0181

Current Biotechnology ›› 2023, Vol. 13 ›› Issue (2): 305-310.DOI: 10.19586/j.2095-2341.2022.0181

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Effects of NRG4 on Proliferation, Apoptosis and Cell Cycle of Acute Myeloid Leukemia Cells

Wanling HUANG¹^,²(), Wenqi ZHU¹^,², Nini GUO¹^,², Nan WANG¹^,², Qian REN¹^,², Xiaotong MA¹^,²()

^1.State Key Laboratory of Experimental Hematology，National Clinical Research Center for Blood Diseases，Haihe Laboratory of Cell Ecosystem，Institute of Hematology & Blood Diseases Hospital，Chinese Academy of Medical Sciences & Peking Union Medical College，Tianjin 300020，China
^2.Tianjin Institutes of Health Science，Tianjin 3016000，China

Received:2022-11-04 Accepted:2022-12-16 Online:2023-03-25 Published:2023-04-07
Contact: Xiaotong MA

Abstract

Abstract:

The aim of the study is to detect the expression of NRG4 in patients with hematopoietic malignancies and malignant hematopoietic cell lines and to investigate its effect on proliferation， apoptosis and clone formation of acute myeloid leukemia （AML） cells．The expression of NRG4 in patients with hematopoietic malignancies and its correlation with the prognosis of patients were analyzed with gene expression omnibus （GEO） and OncoLnc database. Expression of NRG4 mRNA of malignant hematopoietic cell lines were detected by real-time fluorescent quantitative PCR. Lentiviral vector for knockdown of NRG4 were constructed and transduced into AML cell lines Molm-13， KG-1a and Kasumi．The roles of NRG4 in cell proliferation， apoptosis， cell cycle， colony formation were evaluated by cell counting， flow cytometric analysis of Annexin V/7-AAD staining， PI staining， colony-forming assay， respectively. Expression of NRG4 was significantly higher in bone marrow mononuclear cells （BMMCs） from patients with AML compared to those from healthy donors. The survival of AML patients with higher NRG4 expression were significantly shorter than those of AML patients with low NRG4 level（P<0.05）. Knockdown of NRG4 inhibited cell growth and colony formation， promoted apoptosis， and induced cell cycle arrest（P<0.05）. In summary， NRG4 is highly expressed in BMMCs from patients with AML and AML cell lines， and its expression was correlated with poor prognosis in AML patients. NRG4 exerted tumor-promoting roles in AML cells. These findings shed insight into the underlying mechanism of development of AML and provide novel target for AML treatment.

Key words: NRG4 gene, acute myeloid leukemia, cell proliferation, apoptosis

CLC Number:

R733.71

Wanling HUANG, Wenqi ZHU, Nini GUO, Nan WANG, Qian REN, Xiaotong MA. Effects of NRG4 on Proliferation, Apoptosis and Cell Cycle of Acute Myeloid Leukemia Cells[J]. Current Biotechnology, 2023, 13(2): 305-310.

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https://swjsjz.magtechjournal.com/EN/Y2023/V13/I2/305

Figures/Tables 8

Table 1 Primer sequences used in this study

基因		引物序列（5'→3'）
shNRG4	F	5'-CCGGACCTGGTAGAGACGAGCAGTACTCGAGTACTGCTCGTCTCTACCAGGTTTTTTG-3'
shNRG4	R	5'-AATTCAAAAAACCTGGTAGAGACGAGCAGTACTCGAGTACTGCTCGTCTCTACCAGGT-3'
NRG4^*	F	5'-ATGCCAACAGATCACGAAGAG-3'
NRG4^*	R	5'-AATGGGCTGGGAATAGTAGGT-3'
ACTIN^*	F	5'-AGTATCCATGAAAATAAGTGGTTACAGG-3'
ACTIN^*	R	5'-TTTGCAGTACATAATTTACACAGAAGC-3'

Fig. 1 Relative mRNA expression of NRG4 in AML patients and hematopoietic malignant cell lines

Fig. 2 Correlational relationship of the percentage of survival of AML patients stratified by NRG4 expression

Fig. 3 RT-PCR evaluation of NRG4 expression in Molm-13,KG-1a and Kasumi cells with NRG4 knockdown

Fig. 4 The effects of knockdown of NRG4 on proliferation of Molm-13, KG-1a and Kasumi cells

Fig. 5 The effects of knockdown of NRG4 on colony formation of Molm-13, KG-1a and Kasumi cells

Fig. 6 The effects of knockdown of NRG4 on the apoptosis of Molm-13, KG-1a and Kasumi cells

Fig. 7 Effect of NRG4 knockdown on cell cycle of Molm-13, KG-1a and Kasumi cells

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Effects of NRG4 on Proliferation, Apoptosis and Cell Cycle of Acute Myeloid Leukemia Cells

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