朱琳, 陈潞萍, 牟萍. Midkine对神经母细胞瘤TNB1细胞生长与分化的影响[J]. 中国肿瘤临床, 2022, 49(3): 109-114. DOI: 10.12354/j.issn.1000-8179.2022.20211087
引用本文: 朱琳, 陈潞萍, 牟萍. Midkine对神经母细胞瘤TNB1细胞生长与分化的影响[J]. 中国肿瘤临床, 2022, 49(3): 109-114. DOI: 10.12354/j.issn.1000-8179.2022.20211087
shu
Lin Zhu, Luping Chen, Ping Mu. Effect of midkine on cell growth and differentiation of neuroblastoma TNB1 cells[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2022, 49(3): 109-114. DOI: 10.12354/j.issn.1000-8179.2022.20211087
Citation: Lin Zhu, Luping Chen, Ping Mu. Effect of midkine on cell growth and differentiation of neuroblastoma TNB1 cells[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2022, 49(3): 109-114. DOI: 10.12354/j.issn.1000-8179.2022.20211087
shu

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

    摘要
  • 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。

     

    Abstract:
      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.

     

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