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目的:检测Twist-1基因在白血病患者和造血系统恶性肿瘤细胞系中的表达情况,并探讨其高表达对髓系白血病细胞增殖和凋亡的影响。方法:用Real-time PCR检测急性髓系白血病(acute myeloid leukemia,AML)、急性淋巴细胞白血病(acute lymphoid leukemia,ALL)、慢性粒细胞白血病(chronic myeloid leukemia,CML)患者和正常人的骨髓单个核细胞对照以及造血系统恶性肿瘤细胞系中Twist-1 mRNA表达情况。构建Twist-1过表达及干扰载体,制备慢病毒并感染髓系白血病细胞系K562、U937、KG-1a,通过细胞计数实验、集落形成实验、流式细胞术、Annexin V/PI方法评价Twist-1对白血病细胞增殖、集落形成能力、周期、凋亡的影响。结果:Twist-1在AML及CML患者中的表达水平显著高于对照(均P<0.05),而ALL患者与对照组没有显著差异(P>0.05)。在K562、U937、KG-1a中过表达及干扰实验证实,Twist-1高表达促进肿瘤细胞增殖、集落形成并抑制细胞凋亡;干扰Twist-1表达则效果相反。结论:Twist-1高表达于AML、CML髓系白血病细胞,并促进白血病细胞的增殖和抑制凋亡。
OBJECTIVE: To detect the expression of Twist-1 gene in leukemia and hematopoietic malignant tumor cell lines and to investigate the effect of its high expression on the proliferation and apoptosis of myeloid leukemia cells. Methods: Real-time PCR was used to detect single myeloid leukemia (AML), acute lymphocytic leukemia (ALL), chronic myeloid leukemia (CML) Nuclear cell control and hematopoietic malignant tumor cell line Twist-1 mRNA expression. Twist-1 overexpression and interference vectors were constructed, lentivirus was infected and infected with myeloid leukemia cell lines K562, U937 and KG-1a. Twist-1 expression was detected by cell counting, colony formation assay, flow cytometry and Annexin V / 1 on leukemic cell proliferation, colony formation ability, cycle, apoptosis. Results: The expression of Twist-1 in AML and CML patients was significantly higher than that in controls (all P <0.05), while there was no significant difference between ALL patients and controls (P> 0.05). Overexpression and interference experiments in K562, U937 and KG-1a confirmed that Twist-1 overexpression promoted tumor cell proliferation, colony formation and apoptosis, while interfering with Twist-1 expression had the opposite effect. Conclusion: Twist-1 is highly expressed in AML and CML myeloid leukemia cells, and promotes the proliferation and inhibits the apoptosis of leukemia cells.