NRG4基因对急性髓系白血病细胞增殖、凋亡及周期的影响

Effects of NRG4 on Proliferation,Apoptosis and Cell Cycle of Acute Myeloid Leukemia Cells

为了分析、检测造血系统恶性肿瘤患者和细胞系中NRG4基因的表达情况,探讨其对急性髓系白血病(acute myeloid leukemia,AML)细胞系增殖、凋亡、周期和集落形成的影响。运用基因表达综合数据库(gene expression omnibus,GEO)和OncoLnc数据库分析NRG4在造血系统恶性肿瘤患者中的表达情况及其对患者预后的影响,实时荧光定量PCR检测NRG4在造血系统恶性肿瘤各细胞系中的表达情况。构建包含NRG4 shRNA及scramble对照慢病毒,感染AML细胞系Molm-13、KG-1a和Kasumi,采用生长曲线、AnnexinV/7-AAD和PI染色、集落形成等方法评价NRG4对AML细胞增殖、凋亡、周期和集落形成的影响。结果表明,NRG4在AML患者骨髓单核细胞(bone marrow mononuclear cells,BMMCs)中的表达水平显著高于健康对照,高表达NRG4的白血病患者的生存率显著低于低表达患者(P<0.05)。在Molm-13、KG-1a和Kasumi细胞中敲降NRG4能够显著抑制细胞的增殖、集落形成,促进细胞凋亡,并使细胞阻滞于S期(P<0.05)。总之,NRG4基因在AML患者和细胞系中高表达,并与患者预后不良相关,对AML细胞系具有显著的促癌作用。研究结果有助于阐明AML的发生机制,为新治疗靶点的开发奠定了基础。

The aim of the study is to detect the expression of NRG4 in patients with hematopoietic malignancies and malignant hematopoietic cell lines and to investigate its effect on proliferation,apoptosis and clone formation of acute myeloid leukemia(AML)cells.The expression of NRG4 in patients with hematopoietic malignancies and its correlation with the prognosis of patients were analyzed with gene expression omnibus(GEO)and OncoLnc database.Expression of NRG4 mRNA of malignant hematopoietic cell lines were detected by real-time fluorescent quantitative PCR.Lentiviral vector for knockdown of NRG4 were constructed and transduced into AML cell lines Molm-13,KG-1a and Kasumi.The roles of NRG4 in cell proliferation,apoptosis,cell cycle,colony formation were evaluated by cell counting,flow cytometric analysis of Annexin V/7-AAD staining,PI staining,colony-forming assay,respectively.Expression of NRG4 was significantly higher in bone marrow mononuclear cells(BMMCs)from patients with AML compared to those from healthy donors.The survival of AML patients with higher NRG4 expression were significantly shorter than those of AML patients with low NRG4 level(P<0.05).Knockdown of NRG4 inhibited cell growth and colony formation,promoted apoptosis,and induced cell cycle arrest(P<0.05).In summary,NRG4 is highly expressed in BMMCs from patients with AML and AML cell lines,and its expression was correlated with poor prognosis in AML patients.NRG4 exerted tumor-promoting roles in AML cells.These findings shed insight into the underlying mechanism of development of AML and provide novel target for AML treatment.