摘要
目的探讨Septin-9在急性淋巴细胞白血病(ALL)病儿骨髓组织中的表达及对人Jurkat细胞生物学功能的影响。方法选取2014年2月—2017年8月在我院门诊就诊或住院治疗的ALL病儿94例,其中初诊病儿63例,缓解病儿20例,复发病儿11例。选取同期行骨骼矫治手术的病儿16例作为对照组。采用实时荧光定量PCR技术检测骨髓单个核细胞中Septin-9mRNA的表达;培养人Jurkat细胞并分为Septin-9干扰组(C组)、阴性对照组(B组)和空白组(A组),采用Western blot方法检测细胞中Septin-9蛋白表达;采用MTT法检测细胞增殖能力;采用流式细胞术检测细胞周期;采用划痕实验检测细胞迁移能力;采用Transwell小室检测细胞侵袭能力。结果初诊病儿骨髓单个核细胞中Septin-9mRNA相对表达量高于对照组,差异具有统计学意义(t=7.283,P<0.05);初诊病儿中,B-ALL和T-ALL病儿骨髓单个核细胞中Septin-9mRNA相对表达量差异无统计学意义(t=0.812,P>0.05);标危组、中危组和高危组病儿骨髓单个核细胞中Septin-9mRNA相对表达量比较差异均有统计学意义(F=108.770,P<0.05);初诊病儿、缓解病儿和复发病儿骨髓单个核细胞中Septin-9mRNA相对表达量比较差异均有统计学意义(F=51.456,P<0.05);C组细胞中Septin-9蛋白相对表达量及24、48、72和96h细胞吸光度值均低于B组和A组(F=7.349～62.232,P<0.05);C组G_0/G_1期细胞比例高于B组和A组,而S期细胞比例低于B组和A组(F=7.887、8.148,P<0.05);C组24和48h划痕愈合率以及侵袭细胞数均低于B组和A组(F=42.444～97.442,P<0.05)。结论 Septin-9在ALL病儿骨髓单个核细胞中呈高表达,特异性抑制人Jurkat细胞中Septin-9mRNA表达,可抑制细胞迁移和侵袭能力,可能通过改变细胞周期而抑制细胞增殖。
Objective To investigate the expression of Septin-9 in the bone marrow tissue of children with acute lymphoblastic leukemia (ALL) and its effect on the biological function of human Jurkat cells.Methods A total of 94 children with ALL who were treated at the outpatient service or were hospitalized in our hospital from February 2014 to August 2017 were enrolled,among whom 63 were diagnosed for the first time,20 had remission,and 11 had recurrence.A total of 16 patients who underwent orthopedic surgery were enrolled as control group.Quantitative real-time PCR was used to measure the mRNA expression of Septin-9 in bone marrow mononuclear cells.Human Jurkat cells were cultured and divided into blank group (group A),negative control group (group B),and Septin-9 interference group (group C),and Western blot was used to measure the protein expression of Septin-9 in these cells.MTT assay was used to assess cell proliferation,flow cytometry was used to determine cell cycle,wound healing assay was used to evaluate cell migration ability,and Transwell chamber was used to evaluate cell invasiveness.Results The group of newly diagnosed children had significantly higher relative mRNA expression of Septin-9 in bone marrow mononuclear cells than the control group (t=7.283,P<0.05).In the newly diagnosed children,there was no significant difference in the relative mRNA expression of Septin-9 in bone marrow mononuclear cells between the children with B-lineage ALL and those with T-cell ALL (t=0.812,P>0.05).There was a significant difference in the relative mRNA expression of Septin-9 in bone marrow mononuclear cells between the standard risk group,the intermediate risk group,and the high risk group (F=108.770,P<0.05).There was also a significant difference in the relative mRNA expression of Septin-9 in bone marrow mononuclear cells between the newly diagnosed children,the children with remission,and the children with recurrence (F=51.456,P<0.05).Compared with groups A and B,group C had significantly lower relative protein expression of Septin-9 and absorbance values at 24,48,72,and 96 hours (F= 7.349-62.232,P<0.05),a significantly higher proportion of cells in G 0/G 1 phase and a significantly lower proportion of cells in S phase (F=7.887,8.148,P<0.05),and significantly lower wound healing rates at 24 and 48 hours and number of invasive cells (F=42.444-97.442,P<0.05).ConclusionSeptin-9 is highly expressed in bone marrow mononuclear cells in children with ALL.Specific inhibition of Septin-9 mRNA expression in human Jurkat cells can inhibit cell migration and invasion and inhibit cell proliferation by altering cell cycle.
作者
汪玉芳
柯善栋
郑芳
WANG Yufang;KE Shandong;ZHENG Fang(Department of Hematology, Edong Medical Group Huangshi Central Hospital (Hospital Affiliated to Hubei Polytechnic University), Huangshi 435000, China)
出处
《精准医学杂志》
2018年第6期499-504,共6页
Journal of Precision Medicine
基金
湖北省卫生厅青年科技人才项目(QJX2012-06)
