Original Article

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Blood Res 2023; 58(3):

Published online September 30, 2023

https://doi.org/10.5045/br.2023.2023097

© The Korean Society of Hematology

MicroRNA-765 is upregulated in myelodysplastic syndromes and induces apoptosis via PLP2 inhibition in leukemia cells

Seong-Ho Kang1, Ji Seon Choi2

Department of Laboratory Medicine, 1Chosun University College of Medicine, Gwangju, 2International St. Mary’s Hospital, Catholic Kwandong University, Incheon, Korea

Correspondence to : Seong-Ho Kang, M.D., Ph.D.
Department of Laboratory Medicine, Chosun University College of Medicine, 365 Pilmun-daero, Dong-gu, Gwangju 61453, Korea
E-mail: seonghomed@hanmail.net

*This study was supported by a grant from the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Korea government (MSIT) (NRF-2021R1F1A1045955).

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Background
Epigenetic studies, particularly research on microRNA (miRNA), have flourished. The abnormal expression of miRNA contributes to the development of hematologic malignancies. miR-765 has been reported to inhibit cell proliferation by downregulating proteolipid protein 2 (PLP2), which causes apoptosis. We investigated miR-765 dysregulation in myelodysplastic syndromes (MDS).
Methods
We compared the expression profiles of miR-765 in 65 patients with MDS and 11 controls. Cell proliferation and apoptosis assays were performed to determine the in vitro effects of miR-765 on leukemia cells transfected with the miR-765 mimic. Reverse transcription quantitative PCR (RT-qPCR) and western blotting were performed to examine the targets of miR-765.
Results
We found that miR-765 levels were upregulated 10.2-fold in patients with MDS compared to controls. In refractory cytopenia with multilineage dysplasia, the percentage of patients with elevated miR-765 levels was significantly higher than in other forms of MDS. Experiments with leukemia cells revealed that transfection with a miR-765 mimic inhibited cell proliferation and induced apoptosis. RT-qPCR and western blotting demonstrated that the target of miR-765 was PLP2.
Conclusion
These findings imply that upregulation of miR-765 induces apoptosis via downregulation of PLP2 and may have a role in MDS pathogenesis.


Keywords: MDS, microRNA, Deregulation, Apoptosis, Pathogenesis

Article

Original Article

Blood Res 2023; 58(3): 133-137

Published online September 30, 2023 https://doi.org/10.5045/br.2023.2023097

Copyright © The Korean Society of Hematology.

MicroRNA-765 is upregulated in myelodysplastic syndromes and induces apoptosis via PLP2 inhibition in leukemia cells

Seong-Ho Kang1, Ji Seon Choi2

Department of Laboratory Medicine, 1Chosun University College of Medicine, Gwangju, 2International St. Mary’s Hospital, Catholic Kwandong University, Incheon, Korea

Correspondence to:Seong-Ho Kang, M.D., Ph.D.
Department of Laboratory Medicine, Chosun University College of Medicine, 365 Pilmun-daero, Dong-gu, Gwangju 61453, Korea
E-mail: seonghomed@hanmail.net

*This study was supported by a grant from the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Korea government (MSIT) (NRF-2021R1F1A1045955).

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background
Epigenetic studies, particularly research on microRNA (miRNA), have flourished. The abnormal expression of miRNA contributes to the development of hematologic malignancies. miR-765 has been reported to inhibit cell proliferation by downregulating proteolipid protein 2 (PLP2), which causes apoptosis. We investigated miR-765 dysregulation in myelodysplastic syndromes (MDS).
Methods
We compared the expression profiles of miR-765 in 65 patients with MDS and 11 controls. Cell proliferation and apoptosis assays were performed to determine the in vitro effects of miR-765 on leukemia cells transfected with the miR-765 mimic. Reverse transcription quantitative PCR (RT-qPCR) and western blotting were performed to examine the targets of miR-765.
Results
We found that miR-765 levels were upregulated 10.2-fold in patients with MDS compared to controls. In refractory cytopenia with multilineage dysplasia, the percentage of patients with elevated miR-765 levels was significantly higher than in other forms of MDS. Experiments with leukemia cells revealed that transfection with a miR-765 mimic inhibited cell proliferation and induced apoptosis. RT-qPCR and western blotting demonstrated that the target of miR-765 was PLP2.
Conclusion
These findings imply that upregulation of miR-765 induces apoptosis via downregulation of PLP2 and may have a role in MDS pathogenesis.

Keywords: MDS, microRNA, Deregulation, Apoptosis, Pathogenesis

Fig 1.

Figure 1.Relative expression of miR-765 in patients (A) with MDS and controls and (B) with and without trisomy 1q.
Blood Research 2023; 58: 133-137https://doi.org/10.5045/br.2023.2023097

Fig 2.

Figure 2.Cell proliferation after transfection with a miR-765 mimic (miR-765) or a negative control miRNA.
Blood Research 2023; 58: 133-137https://doi.org/10.5045/br.2023.2023097

Fig 3.

Figure 3.(A, B) Annexin V (FITC)/PI assay and (C) caspase assay after transfection with a negative control miRNA (NC) and miR-765 mimic.
Blood Research 2023; 58: 133-137https://doi.org/10.5045/br.2023.2023097

Fig 4.

Figure 4.(A) mRNA analysis of PLP2 and (B) western blot analysis after transfection with a miR-765 mimic (miR-765) or negative control miRNA (NC).
Blood Research 2023; 58: 133-137https://doi.org/10.5045/br.2023.2023097

Patients distribution according to miR-765 expression and MDS subtypes..


MDS subtypesN of patients (%)
Low miR-765 expressionHigh miR-765 expression
RCUD7 (21.9%)2 (6.1%)
RARS0 (0%)1 (3.0%)
RCMD8 (25%)19 (57.6%)
RAEB-15 (16.7%)4 (12.1%)
RAEB-24 (12.5%)6 (1.1%)
MDS U8 (25%)1 (3.0%)
Total32 (100%)33 (100%)

Abbreviations: MDS, myelodysplastic syndromes; MDS U, unclassified myelodysplastic syndromes; RAEB-1, refractory anemia with excess blasts type 1; RAEB-2, refractory anemia with excess blasts type 2; RARS, refractory anemia with ring sideroblasts; RCUD, refractory cytopenia with unilineage dysplasia..



Patients distribution according to miR-765 expression and IPSS-R subgroups..


Prognostic subgroupsN of patients (%)
Low miR-765 expressionHigh miR-765 expression
Very low2 (6.7%)2 (6.7%)
Low4 (13.3%)6 (16.7%)
Intermediate12 (40.0%)10 (33.3%)
High5 (16.7%)4 (13.3%)
Very high7 (23.3%)5 (16.7%)
Total30 (100%)30 (100%)

Abbreviation: IPSS-R, International Prognostic Scoring System..


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