Man Jian-Cheng, Cheng Juan, Zhao Li
The First Clinical Medical College of Lanzhou University; Department of Central Laboratory, The Frist Hospital of Lanzhou University; Gansu Key Laboratory of Genetic Study of Hematopathy, Lanzhou 730000, Gansu Province, China.
The First Clinical Medical College of Lanzhou University; Department of Central Laboratory, The Frist Hospital of Lanzhou University; Gansu Key Laboratory of Genetic Study of Hematopathy, Lanzhou 730000, Gansu Province, China.E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2024 Feb;32(1):52-56. doi: 10.19746/j.cnki.issn.1009-2137.2024.01.009.
To construct a acute myeloid leukemia (AML) cell line in which gene is stably knocked out by CRISPR-Cas9-mediated gene editing technique, so as to clarify the effect of gene knockout on the proliferation of AML cells, and preliminarily explore the role of gene in the pathogenesis of AML.
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 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.
Compared with non-tumor hematological patients, the levels of gene and protein in BMMC of newly diagnosed AML patients were significantly increased ( <0.05). The stable knockout cell line can be obtained by CRISPR-Cas9-Mediated gene editing technique, and the proliferation ability of THP-1 cells with gene knockout was significantly decreased ( <0.05).
is highly expressed in AML cells, and knocking out can significantly affect the proliferation ability of AML cells, which provides a new potential therapeutic target for the precise treatment of AML.
利用CRISPR-Cas9介导的基因编辑技术构建一个基因被稳定敲除的急性髓系白血病（AML）细胞系，以阐明该基因敲除对AML细胞增殖的影响，并初步探讨该基因在AML发病机制中的作用。
分别采用定量实时PCR（qRT-PCR）和蛋白质免疫印迹法检测非肿瘤血液病患者及新诊断AML患者骨髓单个核细胞（BMMC）中HOXA5的表达。利用CRISPR-Cas9介导的基因编辑技术构建基因敲除的AML细胞系KO-HOXA5-THP-1，通过qRT-PCR和蛋白质免疫印迹法验证HOXA5基因的敲除情况，采用CCK-8法检测细胞增殖。
与非肿瘤血液病患者相比，新诊断AML患者BMMC中该基因及蛋白水平显著升高（<0.05）。通过CRISPR-Cas9介导的基因编辑技术可获得稳定的基因敲除细胞系，基因敲除的THP-1细胞增殖能力显著降低（<0.05）。
该基因在AML细胞中高表达，敲除该基因可显著影响AML细胞的增殖能力，为AML的精准治疗提供了新的潜在治疗靶点。
