Abstract
BackgroundAs a disease of hematopoietic stem cell, chronic myeloid leukemia (CML) possesses unique biological and clinical features. However, the biologic mechanism underlying its development remains poorly understood. Thus, the objective of the present study is to discuss the effect of cytidine deaminase (CDA) gene silencing on the apoptosis and proliferation of CML K562 cells.MethodsCDA mRNA expression was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and enzymatic activity of CDA was measured by a nuclide liquid scintillation method. RT-qPCR and Western blot analysis were used to detect CDA mRNA and protein expression. Cell proliferation, apoptosis and cell cycle were measured by CCK-8 assay and flow cytometry. The expression of proteins relevant to cell proliferation, apoptosis and cell cycle was measured by Western blot analysis. Tumor xenografts were implanted in nude mice to verify the effect of CDA silencing on tumor growth in vivo.ResultsCML and AL patients showed increased mRNA expression and enzymatic activity of CDA. Compared with the blank group, the mRNA and protein expression of CDA in the shRNA-1 and shRNA-2 groups decreased significantly. As a result, the proliferation of K562 cells was inhibited after CDA silencing and the cells were mainly arrested in S and G2 phases, while the apoptosis rate of these cells was increased. In addition, CDA gene silencing in K562 cells led to down-regulated p-ERK1/2, t-AKT, p-AKT and BCL-2 expression and up-regulated expression of P21, Bax, cleaved caspase-3/total caspase-3 and cleaved PARP/total PARP. Finally, CDA gene silencing inhibited tumor growth.ConclusionOur study demonstrated that CDA gene silencing could inhibit CML cell proliferation and induce cell apoptosis. Therefore, CDA gene silencing may become an effective target for the treatment of leukemia.
Highlights
As a disease of hematopoietic stem cell, chronic myeloid leukemia (CML) possesses unique biological and clinical features
Chronic myeloid leukemia (CML) is a hematopoietic malignant disease characterized by the presence of the Philadelphia chromosome (Ph) due to the reciprocal translocation of chromosomes 9 and 22, resulting in the formation of a BCR–ABL fusion onco-protein [1,2,3]
cytidine deaminase (CDA) mRNA and protein expression was increased in CML and acute leukemia (AL) patients reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to detect CDA mRNA expression in bone marrow mononuclear cells of 112 CML and 106 AL patients, as well as in 101 donors of hematopoietic stem cell transplantation
Summary
As a disease of hematopoietic stem cell, chronic myeloid leukemia (CML) possesses unique biological and clinical features. Chronic myeloid leukemia (CML) is a hematopoietic malignant disease characterized by the presence of the Philadelphia chromosome (Ph) due to the reciprocal translocation of chromosomes 9 and 22, resulting in the formation of a BCR–ABL fusion onco-protein [1,2,3]. As a myelo-proliferative neoplasm, the incidence of CML is higher than 2/100,000 [4]. The incidence of CML in females is lower than that in males [5]. The BCR– ABL fusion protein leads to the pathogenicity of CML, while the therapies targeting the BCR–ABL protein have encountered many difficulties [6]. Gene therapy has become more common recently in the treatment of CML.
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