ROBO3在儿童急性髓系白血病患者中的表达及其对细胞增殖和凋亡的影响

Expression of ROBO3 and Its Effect on Cell Proliferation and Apoptosis in Pediatric Patients with Acute Myeloid Leukemia

目的:研究ROBO3在儿童急性髓系白血病(AML)患者中的表达及其对AML细胞增殖和凋亡的影响。方法:采用荧光定量PCR法检测不同治疗阶段的儿童AML患者骨髓中ROBO3的表达,分析初诊患儿ROBO3表达量与临床特征的关系。同时采用电转染的方法靶向干扰ROBO3表达,应用CCK-8法检测其对AML细胞系(HL-60和THP-1)增殖的影响,流式细胞术检测其对凋亡的作用。结果:与健康对照组相比,ROBO3在初诊儿童AML患者中显著高表达,尤其是在非M3型、年龄偏小的患者(<10岁)及高危组中高表达。而且与初发组相比,治疗后完全缓解组ROBO3 mRNA的表达水平显著下降。靶向ROBO3可以有效抑制AML细胞的增殖,并促进细胞凋亡。结论:ROBO3在儿童AML患者中差异表达,很可能是儿童AML潜在的标志物及新靶点。

Objective:To investigate the expression of ROBO3 in pediatric AML patients and explore its function on cell proliferation and apoptosis.Methods:The expression of ROBO3 in pediatric AML patients at different treatment stage was detected by real-time quantitative polymerase chain reaction(RT-q PCR).The relationship between the expression of ROBO3and clinic pathological characteristics in newly diagnosed pediatric AML patients was analyzed.Moreover,the effects of ROBO3 on the proliferation and apoptosis of AML cell lines HL-60 and THP-1 were estimated by using CCK-8 and flow cytometry after transfection with ROBO3 siRNA.Results:It was found that ROBO3 expression was significantly increased in most of newly diagnosed pediatric AML patients,especially in non-M3 subtype,younger patients(<10 years old),and high risk group,compared to corresponding controls.Furthermore,the expression level of ROBO3 was sharply decreased in patients who achieved complete remission.Targeting ROBO3 significantly inhibited AML cell proliferation,as well as increased apoptosis by ROBO3 siRNA transfection in vitro.Conclusion:ROBO3 is differentially expressed within distinct subtypes of the pediatric AML patients,which suggested that ROBO3 may be a potential biomarker and a new therapeutic target for pediatric AML.