Midkine对神经母细胞瘤TNB1细胞生长与分化的影响  

Effect of midkine on cell growth and differentiation of neuroblastoma TNB1 cells

作　　者：朱琳 陈潞萍 牟萍 Lin Zhu;Luping Chen;Ping Mu(Department of Biochemistry and Molecular Biology,Shenyang Medical College,Shenyang 110034,China;Department of Physiology,Basic Medical School,Shenyang Medical College,Shenyang 110034,China)

机构地区：[1]沈阳医学院基础医学院生物化学与分子生物学教研室,沈阳市110034 [2]沈阳医学院基础医学院生理学教研室,沈阳市110034

出　　处：《中国肿瘤临床》2022年第3期109-114,共6页Chinese Journal of Clinical Oncology

基　　金：辽宁省“兴辽英才计划”青年拔尖人才项目(编号:XLYC1807116);辽宁省自然科学基金项目(编号:20180550491)资助。

摘　　要：目的:通过对中期因子(midkine,MK)在神经母细胞瘤TNB1细胞中的生物学作用进行研究,揭示该因子在肿瘤细胞生长和分化中相关分子机制。方法:1)利用神经母细胞瘤临床患者数据库,明确MK的表达与神经母细胞瘤患者生存率之间的关系。在体外细胞实验中,利用携带MK shRNA载体的慢病毒体外感染神经母细胞瘤TNB1细胞,分析MK的敲减对其生长与分化的影响;2)通过进一步临床数据分析,找到与MK表达的相关基因,并采用Real-time PCR验证MK对相关基因之间表达联系;3)联合使用携带MK shRNA和肾细胞癌下调蛋白1(downregulated in renal carcinoma 1,DRR1)shRNA载体的两种慢病毒感染TNB1细胞,评价MK-DRR1轴对TNB1细胞分化以及克隆形成的影响。结果:临床数据分析显示MK的表达量与神经母细胞瘤患者生存率呈负相关(P<0.01)。体外细胞实验结果表明MK的敲减能够抑制TNB1细胞生长,同时促进TNB1细胞分化;临床数据分析结果进一步证明,在神经母细胞瘤中MK与DRR1的表达量呈负相关(P<0.01);Real-time PCR结果证实下调MK表达可增强TNB1细胞中DRR1的表达(P<0.01);细胞挽救实验显示敲低DRR1的表达可以挽救由MK敲减而产生的对TNB1细胞的分化和软琼脂克隆的影响(P<0.01)。结论:MK通过调控DRR1的表达从而参与调节TNB1细胞的生长与分化。Objective:To assess biological functions and underlying mechanisms of midkine(MK)in cell growth and differentiation of neuroblastoma TNB1 cells.Methods:First,the correlation between MK expression and survival rate was assessed in patients with neuroblastoma whose data were obtained from a clinical database.Furthermore,cell growth and differentiation ability of TNB1 cells were evaluated after treatment with MK shRNA-containing lentivirus in vitro.Second,the related gene of MK was screened out by analyzing data from the clinical database,and its expression in MK shRNA-treated TNB1 cells was assessed using real-time PCR.Finally,TNB1 cells were infected with MK shRNA-and downregulated in renal carcinoma 1(DRR1)shRNA-containing lentivirus,and the effect of MK-DRR1 axis on cell differentiation and soft agar colony formation was evaluated.Results:Data from the clinical database revealed the negative correlation between MK expression and survival rate of patients with neuroblastoma(P<0.01).Downregulation of MK repressed cell growth and promoted TNB1 cell differentiation.Analysis of the clinical database revealed a negative correlation between MK expression and DRR1 expression(P<0.01).Real-time PCR results confirmed that DRR1 expression was increased in MK shRNA-treated TNB1 cells(P<0.01).Combined treatment with MK shRNA and DRR1 shRNA rescued the effect of MK shRNA on cell differentiation and soft agar colony formation(P<0.01).Conclusions:MK mediated the growth and differentiation of TNB1 cells by regulating DRR1 expression.

关 键 词：中期因子 神经母细胞瘤 生长 分化 肾细胞癌下调蛋白1 

分 类 号：R739.4[医药卫生—肿瘤]