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原代培养人喉鳞癌细胞中SIX1、TGF-β、VEGF-C表达的相关性

张继华, 陈春菊, 韩彩莉, 苏静, 李丹, 王来

张继华, 陈春菊, 韩彩莉, 苏静, 李丹, 王来. 原代培养人喉鳞癌细胞中SIX1、TGF-β、VEGF-C表达的相关性[J]. 肿瘤防治研究, 2016, 43(6): 453-458. DOI: 10.3971/j.issn.1000-8578.2016.06.004
引用本文: 张继华, 陈春菊, 韩彩莉, 苏静, 李丹, 王来. 原代培养人喉鳞癌细胞中SIX1、TGF-β、VEGF-C表达的相关性[J]. 肿瘤防治研究, 2016, 43(6): 453-458. DOI: 10.3971/j.issn.1000-8578.2016.06.004
ZHANG Jihua, CHEN Chunju, HAN Caili, SU Jing, LI Dan, WANG Lai. Correlation Between Expression of SIX1, TGF-β and VEGF-C in Primarily Cultured Human Laryngeal Squamous Cell Carcinoma Cells[J]. Cancer Research on Prevention and Treatment, 2016, 43(6): 453-458. DOI: 10.3971/j.issn.1000-8578.2016.06.004
Citation: ZHANG Jihua, CHEN Chunju, HAN Caili, SU Jing, LI Dan, WANG Lai. Correlation Between Expression of SIX1, TGF-β and VEGF-C in Primarily Cultured Human Laryngeal Squamous Cell Carcinoma Cells[J]. Cancer Research on Prevention and Treatment, 2016, 43(6): 453-458. DOI: 10.3971/j.issn.1000-8578.2016.06.004

原代培养人喉鳞癌细胞中SIX1、TGF-β、VEGF-C表达的相关性

详细信息
    作者简介:

    张继华(1978-),男,硕士,主治医师,主要从事头颈肿瘤的基础和临床研究

    通讯作者:

    王来,E-mail: wanglai21@126.com

  • 中图分类号: R739.65

Correlation Between Expression of SIX1, TGF-β and VEGF-C in Primarily Cultured Human Laryngeal Squamous Cell Carcinoma Cells

More Information
  • 摘要:
    目的 

    探讨原代人喉鳞癌细胞中SIX1、TGF-β、VEGF-C表达相关性。

    方法 

    取新鲜人喉鳞状细胞癌组织,进行原代细胞培养,利用基因干扰技术,制备SIX1、TGF-β及SIX1+TGF-β靶向siRNA转染至人喉鳞癌细胞,转染成功后将实验细胞分为5组: A组(未转染组)、B组(转染空载体组)、C组(SIX1-siRNA组)、D组(TGFβ-siRNA组)、E组(SIX1+TGF-β-siRNA组)。Western blot及细胞爬片免疫荧光检测各组细胞SIX1、TGF-β及VEGF-C蛋白的表达。RT-PCR检测各组细胞SIX1、TGF-β及VEGF-C mRNA的表达。

    结果 

    SIX1、TGF-β、VEGF-C蛋白及其mRNA的表达在未转染组、转染空载体组中差异无统计学意义。同未转染组相比,SIX1蛋白及其mRNA的表达在SIX1-siRNA组降低的同时出现了VEGF-C表达的降低;TGF-β蛋白及其mRNA的表达在TGFβ-siRNA组降低的同时也出现了VEGF-C表达的降低,差异均有统计学意义。同SIX1-siRNA组、 TGFβ-siRNA组相比,VEGF-C蛋白及其mRNA的表达在SIX1+TGF-β-siRNA组未出现进一步降低。

    结论 

    SIX1和TGF-β能够共同作用促进VEGF-C蛋白及其 mRNA的表达,进而促进人喉鳞癌的淋巴转移。SIX1、TGF-β的变化可能成为临床抑制人喉鳞癌淋巴结转移的潜在靶点。

     

    Abstract:
    Objective 

    To investigate the relationship between sine oculis homeobox homolog 1(SIX1) , TGF-β and VEGF-C in primarily cultured human laryngeal squamous cell carcinoma cells.

    Methods 

    We took fresh human laryngeal squamous cell carcinoma tissues and cultured the primary cells. The technology of RNA interference was used for preparing SIX1-targeting, TGF-β-targeting and SIX1+TGF-β-targeting siRNA to transfect laryngeal squamous cell carcinoma cells. After successful transfection, the experiments were divided into five groups: Group A (untransfected), Group B(empty vector), Group C(SIX1-siRNA), Group D(TGF-β-siRNA) and Group E(SIX1+TGF-β-siRNA). The protein expression of SIX1, TGF-β and VEGF-C were determined by immunohistochemistry and Western blot. The mRNA levels of SIX1, TGF-β and VEGF-C were determined by RT-PCR.

    Results 

    The protein and mRNA expression of SIX1, TGF-β and VEGF-C in empty vector group had no significant difference compared with untransfected group. In SIX1- siRNA group, the expression of SIX1 protein and mRNA were decreased, at the same time the expression of VEGF-C was decreased, with statistically significant difference compared with untransfected group. In TGF- β-siRNA group, the expression of TGF-β protein and mRNA were decreased, at the same time the expression of VEGF-C was decreased, with statistically significant difference compared with untransfected group. Compared with SIX1-siRNA and TGF-β-siRNA groups, the expression of VEGF-C protein and mRNA had no significant decrease in SIX1+TGF-β-siRNA group.

    Conclusion 

    SIX1 could promote lymph node metastasis by coordinating with TGF-β to increase the expression of VEGF-C protein and mRNA, suggesting that SIX1 and TGF-β might be potential therapeutic targets for preventing lymph node metastasis from human laryngeal squamous cell carcinoma.

     

  • 图  1   人喉鳞癌细胞中SIX1、TGF-β和VEGF-C蛋白表达 (×200)

    Figure  1   Expressions of SIX1, TGF-β and VEGF-C proteins in human laryngeal squamous cell carcinoma cells (×200)

    图  2   人喉鳞癌细胞中SIX1、TGF-β和VEGF-C mRNA的表达

    Figure  2   Expressions of SIX1, TGF-β and VEGF-C mRNA in human laryngeal squamous cell

    表  1   Western blot结果显示各试验组SIX1、TGF-β和VEGF-C蛋白表达

    Table  1   Expressions of SIX1, TGF-β and VEGF-C proteins in human laryngeal squamous cell carcinoma cells in each experimental groups detected by Western blot

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出版历程
  • 收稿日期:  2015-07-02
  • 修回日期:  2016-01-16
  • 网络出版日期:  2024-02-04
  • 刊出日期:  2016-06-24

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