Bioinformatics Analysis And Verification Of Aging-Related Genes Of Bone Marrow Mesenchymal Stem Cells In Acute Myeloid Leukemia

BackgroundAcute myelocytic leukemia(AML)is the most common disease in blood diseases.Most clinical manifestations of AML are reflected in the accumulation of rare malignant,poorly differentiated bone marrow cells in the bone marrow,peripheral blood and other organs.Although the traditional chemotherapy method can temporarily alleviate the progress of the disease in newly diagnosed patients,it is basically impossible for the relapsed patients to obtain significant effects.We need to find new therapeutic targets and new treatments,and it is a top priority to extend the overall survival time of AML patients.MSC commonly used in clinical cell therapy,is found in patients with AML in BM-MSC appeared in the case of premature aging.The premature aging MSC produces many adverse effects to hematopoietic microenvironment,hematopoietic regulation and hematopoietic reconstitution.The premature aging MSC deteriorates the course of the development of AML and it also affects the efficiency of hematopoietic stem cell transplantation for AML.Therefore,in-depth study of aging of BM-MSC in AML patients has become the primary problem.ObjectivesThis study was investigated(aimed)to screen the differential expression genes related to aging of bone marrow mesenchymal stem cells(BM-MSCs)in acute myeloid leukemia(AML)patients by bioinformatics methods,to predict and validate the target genes,and provide new molecular markers for the clinical treatment and scientific research of BM-MSCs in AML patients.Methods(1)The gene expression profiling chip related to BM-MSCs in AML patients in our hospital and the gene chip GSE84881 selected from NCBI database GEO were used for data analysis and exploration.(2)The DAVID analysis software was used to perform gene ontology(GO)enrichment analysis and KEGG pathway enrichment analysis.Furthermore,the differential expression genes associated with aging of BM-MSCs in AML patients were identified.(3)Bone marrow samples were collected and MSCs were amplified in vitro and RT-PCR was used to verify the differential expression genes that were further identified with senescence-associated β-galactosidase dyeing and MTT cell proliferation assays.ResultsA total of 247 differential expression genes were filtrated by bioinformatics methods,including 132 up-go genes and 115 down-go genes.Six differential expression genes related to aging of BM-MSCs in AML patients were filtrated,including up-go genes,including COL3A1(P=0.025),CRYAB(P=0.0003),DCN(P=0.0368),and down-go genes,including CCL2(P=0.0256),CTSC(P=0.0001)and IL6(P=0.0482).These six differential expression genes were consistent with chip trends and had a significant correlation with aging of BM-MSCs in AML patients.Meanwhile,the positive rate of senescence-associated(3-galactosidase dyeing in BM-MSCs of AML patients was significantly different from that of healthy donors(P<0.0001).MTT cell proliferation assay showed that BM-MSCs in AML patients had worse proliferative ability than healthy donors’ BM-MSCs.ConclusionThe data here suggest novel clues for the scientific research and clinical treatment of BM-MSCs aging in AML patients.