| Purpose:Acute myeloid leukemia(AML)is a malignant neoplastic disease with abnormal development of the hematopoietic system,also known as blood cancer.The change in epigenetic state is considered to be one of the most important factors to the AML patients.Recent studies have revealed that A-type lamin,expressed by LMNA gene,as an important component protein of cytoskeleton,not only maintains the basic morphology of nucleus,but also plays an important role in epigenetic regulation.However,whether changes in epigenetic state are associated with dysfunction of LMNA in AML are not well understood.Therefore,we need to clarify the expression and molecular mechanism of LMNA in AML,so as to explore the possibility of LMNA as a potential therapeutic target,and to provide a theoretical basis for the elucidation of the pathogenesis of AML.Methods:We analyzed the expression of LMNA gene in the RNA-seq data of AML and healthy people based on GEO and other public databases.Then,the relationship between different expression levels of LMNA in AML and clinical characteristics was analyzed based on UNLCAN and other databases.Based on the Linkedomics website,the LMNA-related genes in AML and the regulatory pathways in which these genes are enriched were further analyzed.The interaction network between LMNA and related genes in AML was analyzed based on Cytoscape software and STRING website.Based on the biosynthesis data,the function of the LMNA gene was further verified at the cell level in vitro.Lentivirus transfection with sh RNA was used to knockdown LMNA in CD34~+hematopoietic stem cells isolated from human umbilical cord blood and induce erythroid differentiation in vitro.The cell proliferation level was analyzed by cell counting method.Flow cytometry was used to detect the early differentiation,terminal differentiation,apoptosis level and enucleation ability of cells.The whole cell and nucleus were observed by Gimsa staining and electron microscopy.Immunofluorescence staining was used to detect the changes of cellular histone markers.Results:(1)The results of expression analysis showed that the expression level of LMNA was different from that of healthy people and AML patients,and higher expression level in AML patients(P<0.05).(2)LMNA gene-related survival analysis showed that high and low expression of LMNA had a significant difference in overall survival,and the high expression group had a lower overall survival rate(P<0.05).(3)The correlation coefficients of LMNA with other genes were obtained.GO enrichment analysis showed that the sets of genes negatively associated with LMNA were mainly located in chromosomal regions,mainly involved in ribosome synthesis,m RNA transcription and DNA repair,and function as histone binding and m RNA binding.KEGG analysis showed that these negatively associated genes were mainly enriched in Fanconi anemia pathway.Protein network construction revealed that the proteins that interact directly with LMNA function primarily in epigenetic regulation.(4)An LMNA knockdown model of cord blood-derived CD34~+cells was successfully established and induced to erythroid differentiation in vitro.The results showed that cell proliferation was inhibited and apoptosis was increased after LMNA knockdown;cell enucleation was inhibited,nuclear morphology was altered and chromatin structure was loosened;and the localization of the histone marker H3K4me3 in the promoter region was altered.Conclusions:(1)LMNA is significantly correlated with epigenetic regulation and Fanconi anemia pathway,we purpose that LMNA is highly associated with dyserythropoiesis in AML and act through epigenetic modifications.(2)Dyserythropoiesis in vitro culture after LMNA knockdown may be caused by altering localization of H3K4me3,which leads to change epigenetic state. |
PDF Full Text Request