Effect of Knocking Out HOXA5 Gene by CRISPR-Cas9-Mediated Gene Editing Technique on Proliferation of Acute Myeloid Leukemia Cells
Objective:To construct a acute myeloid leukemia(AML)cell line in which HOXA5 gene is stably knocked out by CRISPR-Cas9-mediated gene editing technique,so as to clarify the effect of HOXA5 gene knockout on the proliferation of AML cells,and preliminarily explore the role of HOXA5 gene in the pathogenesis of AML.Methods:The expression of HOXA5 in bone marrow mononuclear cells(BMMC)of non-tumor hematological patients and newly diagnosed AML patients was detected by quantitative real-time PCR(qRT-PCR)and Western blot,respectively.The AML cell line KO-HOXA5-THP-1 was constructed in which HOXA5 gene was knocked out by CRISPR-Cas9-Mediated gene editing technique,and the knockout of HOXA5 gene was verified by qRT-PCR and Western blot,and the cell proliferation was detected by CCK-8 assay.Results:Compared with non-tumor hematological patients,the levels of HOXA5 gene and protein in BMMC of newly diagnosed AML patients were significantly increased(P＜0.05).The stable HOXA5 knockout cell line can be obtained by CRISPR-Cas9-Mediated gene editing technique,and the proliferation ability of THP-1 cells with HOXA5 gene knockout was significantly decreased(P＜0.05).Conclusion:HOXA5 is highly expressed in AML cells,and knocking out HOXA5 can significantly affect the proliferation ability of AML cells,which provides a new potential therapeutic target for the precise treatment of AML.
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