PUBLICATION
Targeting lactate dehydrogenase A (LDHA) exerts antileukemic effects on T-cell acute lymphoblastic leukemia
- Authors
- Yu, H., Yin, Y., Yi, Y., Cheng, Z., Kuang, W., Li, R., Zhong, H., Cui, Y., Yuan, L., Gong, F., Wang, Z., Li, H., Peng, H., Zhang, G.
- ID
- ZDB-PUB-200822-19
- Date
- 2020
- Source
- Cancer communications (London, England) 40(10): 501-517 (Journal)
- Registered Authors
- Keywords
- CRISPR/Cas9 gene-editing, LDHA, T-cell lymphoblastic leukemia, oxamate, transgenic zebrafish model
- MeSH Terms
-
- Adult
- Animals
- Animals, Genetically Modified
- Female
- Gene Knockdown Techniques
- PubMed
- 32820611 Full text @ Cancer Commun (Lond)
Abstract
Background T-cell acute lymphoblastic leukemia (T-ALL) is an uncommon and aggressive subtype of acute lymphoblastic leukemia (ALL). In the serum of T-ALL patients, the activity of lactate dehydrogenase A (LDHA) is increased. We proposed that targeting LDHA may be a potential strategy to improve T-ALL outcomes. The current study was conducted to investigate the antileukemic effect of LDHA gene-targeting treatment on T-ALL and the underlying molecular mechanism.
Methods Primary T-ALL cell lines Jurkat and DU528 were treated with the LDH inhibitor oxamate. MTT, colony formation, apoptosis, and cell cycle assays were performed to investigate the effects of oxamate on T-ALL cells. Quantitative real-time PCR (qPCR) and Western blotting analyses were applied to determine the related signaling pathways. A mitochondrial reactive oxygen species (ROS) assay was performed to evaluate ROS production after T-ALL cells were treated with oxamate. A T-ALL transgenic zebrafish model with LDHA gene knockdown was established using CRISPR/Cas9 gene-editing technology, and then TUNEL, Western blotting, and T-ALL tumor progression analyses were conducted to investigate the effects of LDHA gene knockdown on T-ALL transgenic zebrafish.
Results Oxamate significantly inhibited proliferation and induced apoptosis of Jurkat and DU528 cells. It also arrested Jurkat and DU528 cells in G0/G1 phase and stimulated ROS production (all P < 0.001). Blocking LDHA significantly decreased the gene and protein expression of c-Myc, as well as the levels of phosphorylated serine/threonine kinase (AKT) and glycogen synthase kinase 3 beta (GSK-3β) in the phosphatidylinositol 3'-kinase (PI3K) signaling pathway. LDHA gene knockdown delayed disease progression and down-regulated c-Myc mRNA and protein expression in T-ALL transgenic zebrafish.
Conclusion Targeting LDHA exerted an antileukemic effect on T-ALL, representing a potential strategy for T-ALL treatment.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping