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Zwint高表达对肝癌细胞增殖和肝癌肝移植预后的影响

李慧 汪根树 郑俊 蔡建业 张俊斌 程道柔 周奇 杨扬

李慧, 汪根树, 郑俊, 等. Zwint高表达对肝癌细胞增殖和肝癌肝移植预后的影响[J]. 器官移植, 2018, 9(2): 122-129. doi: 10.3969/j.issn.1674-7445.2018.02.006
引用本文: 李慧, 汪根树, 郑俊, 等. Zwint高表达对肝癌细胞增殖和肝癌肝移植预后的影响[J]. 器官移植, 2018, 9(2): 122-129. doi: 10.3969/j.issn.1674-7445.2018.02.006
Li Hui, Wang Genshu, Zheng Jun, et al. Effect of high expression of Zwint on the proliferation of hepatoma cells and the prognosis of liver transplantation for hepatocellular carcinoma[J]. ORGAN TRANSPLANTATION, 2018, 9(2): 122-129. doi: 10.3969/j.issn.1674-7445.2018.02.006
Citation: Li Hui, Wang Genshu, Zheng Jun, et al. Effect of high expression of Zwint on the proliferation of hepatoma cells and the prognosis of liver transplantation for hepatocellular carcinoma[J]. ORGAN TRANSPLANTATION, 2018, 9(2): 122-129. doi: 10.3969/j.issn.1674-7445.2018.02.006

Zwint高表达对肝癌细胞增殖和肝癌肝移植预后的影响

doi: 10.3969/j.issn.1674-7445.2018.02.006
基金项目: 

国家自然科学基金面上项目 81170422

广东省科技计划社会发展领域项目 2017A020215178

广东省科技计划项目 2014A020212717

广州市计划项目科学研究专项 2014J4100128

详细信息
    作者简介:

    李慧,男,1992年生,硕士研究生,研究方向为肝癌的临床与基础研究,Email: lihealth0303@163.com

    通讯作者:

    汪根树,男,1968年生,博士,主任医师,研究方向为肝癌的临床与基础研究,Email: wgsh168@163.com

  • 中图分类号: R617, R735.7

Effect of high expression of Zwint on the proliferation of hepatoma cells and the prognosis of liver transplantation for hepatocellular carcinoma

More Information
  • 摘要:   目的  探讨Zw10结合因子(Zwint)在原发性肝癌(肝癌)中的表达情况及其对肝癌肝移植预后的影响。  方法  收集50例肝癌肝移植受体的肝癌组织、癌旁组织及临床资料。采用实时荧光定量聚合酶链反应(PCR)、蛋白免疫印迹(Western Blot)法和免疫组织化学(免疫组化)法,分别比较20例肝癌肝移植受体的20对肝癌组织及癌旁组织中Zwint信使核糖核酸(mRNA)表达情况和Zwint蛋白的表达水平;选取两个成功干扰Zwint表达的肝癌细胞系HepG-2(si-Zwint-1组和si-Zwint-2组),以空白对照作为si-NC组,采用细胞计数试剂盒(CCK)-8实验、平板克隆实验和细胞周期实验,比较各组细胞的增殖能力和细胞周期;采用Western Blot法和免疫组化法,分析肝癌组织和肝癌细胞中Zwint与细胞周期蛋白D1(cyclin D1)表达的一致性;以Zwint蛋白表达水平的中位数为分割点,将入组病例分高表达组(22例)和低表达组(28例),分析Zwint蛋白表达水平与肝癌肝移植受体临床特征、总体存活率及无复发存活率的关系。  结果  实时荧光定量PCR结果提示肝癌组织中Zwint mRNA表达水平高于癌旁组织(P=0.03)。Western Blot及免疫组化结果均提示肝癌组织中Zwint蛋白的表达水平高于癌旁组织(均为P < 0.05)。干扰肝癌细胞系HepG-2的Zwint基因后,CCK-8及平板克隆实验显示细胞增殖能力明显减弱(均为P < 0.01),细胞周期阻滞于G1期(均为P < 0.05)。Zwint蛋白表达水平与肿瘤直径和肿瘤、淋巴、转移(TNM)分期密切相关(均为P < 0.05)。Zwint高表达组肝癌肝移植受体的总体存活率低于低表达组(P=0.02)。  结论  Zwint高表达于肝癌组织,其通过调节细胞周期促进肝癌细胞增殖,且Zwint表达水平与肝癌肝移植后预后呈负相关。

     

  • 图  1  肝癌组织及癌旁组织Zwint mRNA和Zwint蛋白表达情况

    A图为实时荧光定量PCR分析肝癌组织及其配对的癌旁组织Zwint mRNA表达情况;B图为Western Blot法分析肝癌组织及其配对的癌旁组织Zwint蛋白表达情况,T为肝癌组织,N为癌旁组织;C图为免疫组化法分析肝癌组织及癌旁组织Zwint蛋白表达情况(核内呈褐色的为Zwint阳性细胞,呈蓝色的为Zwint阴性细胞)

    Figure  1.  Expression of Zwint mRNA and Zwint protein in liver cancer tissues and paraneoplastic tissues

    图  2  各组肝癌细胞的增殖能力和细胞周期情况

    A图为验证干扰效率;B图为CCK-8实验分析各组细胞的增殖能力;C图为平板克隆实验分析各组细胞的单细胞成团能力,与si-NC组比较,aP<0.01;D图为细胞周期实验分析各组细胞的细胞周期情况,与si-NC组比较,aP<0.05

    Figure  2.  Cell proliferation and cell cycle of hepatoma carcinoma cells in each group

    图  3  Zwint、cyclin D1及其相关蛋白在肝癌组织和细胞内的表达情况

    Figure  3.  Expression of Zwint, cyclin D1 and correlative protein in liver cancer tissues and cells

    图  4  Zwint高表达组和低表达组的总体生存曲线和无复发生存曲线

    Figure  4.  Total survival curve and recurrence free survival curve of high Zwint expression group and low Zwint expression group

    表  1  Zwint表达与肝癌受体临床特征指标的关系

    Table  1.   Relationship between Zwint expression and clinicopathological characteristics of hepatocellular carcinoma recipients[n(%)]

    指标 n Zwint表达 P
    低表达组(n=28) 高表达组(n=22)
    性别
      男 36 20(40) 16(32) 0.919
      女 14 08(16) 06(12)
    年龄
      ≤55岁 35 21(42) 14(28) 0.384
      > 55岁 15 07(14) 8(16)
    肝功能Child-Pugh分级
      A级 29 19(38) 10(20) 0.278
      B级 16 07(14) 09(18)
      C级 05 02(4)0 3(6)
    甲胎蛋白
      ≤400 μg/L 34 22(44) 12(24) 0.071
      > 400 μg/L 16 6(12) 10(20)
    肿瘤直径
      ≤5 cm 36 26(52) 10(20) < 0.0010
      > 5 cm 14 2(4) 12(24)
    微血管侵犯
      有 09 5(10) 4(8) 0.976
      否 41 23(46) 18(36)
    TNM分期
       < 3期 36 24(48) 12(24) 0.015
      ≥3期 14 4(8) 10(20)
    下载: 导出CSV
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出版历程
  • 收稿日期:  2018-01-03
  • 网络出版日期:  2021-01-19
  • 刊出日期:  2018-03-15

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