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中华口腔医学研究杂志(电子版) ›› 2015, Vol. 09 ›› Issue (01) : 29 -36. doi: 10.3877/cma.j.issn.1674-1366.2015.01.005

所属专题: 文献

基础研究

RNA结合蛋白QKI-5表达对人头颈鳞癌细胞株的影响
马超1,(), 闫威1, 范龙坤1, 郭燕军1   
  1. 1. 061001 沧州市中心医院口腔颌面外科
  • 收稿日期:2014-10-28 出版日期:2015-02-01
  • 通信作者: 马超

Effects of RNA-binding protein quaking 5 on proliferation and cell cycle of head and neck cancer cell lines

Chao Ma1,(), Wei Yan1, Longkun Fan1, Yanjun Guo1   

  1. 1. Oral and Maxillofacial Surgery, Cangzhou Central Hospital, Cangzhou 061001, China
  • Received:2014-10-28 Published:2015-02-01
  • Corresponding author: Chao Ma
  • About author:
    Corresponding author: Ma Chao, Email: , Tel: 0317-2075544
引用本文:

马超, 闫威, 范龙坤, 郭燕军. RNA结合蛋白QKI-5表达对人头颈鳞癌细胞株的影响[J/OL]. 中华口腔医学研究杂志(电子版), 2015, 09(01): 29-36.

Chao Ma, Wei Yan, Longkun Fan, Yanjun Guo. Effects of RNA-binding protein quaking 5 on proliferation and cell cycle of head and neck cancer cell lines[J/OL]. Chinese Journal of Stomatological Research(Electronic Edition), 2015, 09(01): 29-36.

目的

研究QKI-5对人头颈鳞癌细胞株的生长抑制作用。

方法

采用Western blot和免疫组化方法检测人头颈鳞癌临床组织样本中的QKI-5的蛋白表达情况。常规培养舌鳞癌SCC15细胞、舌鳞癌SCC25细胞和头颈鳞癌PCI13细胞,以5 × 104/ml的密度种植细胞。采用Western blot和反转录聚合酶链反应(RT-PCR)检测三种头颈鳞癌细胞株中的QKI-5的蛋白和mRNA表达情况。采用病毒介导QKI-5在SCC25细胞中过表达和在PCI13细胞中沉默QKI-5。采用MTT法和流式细胞法检测SCC25细胞中过表达QKI-5和在PCI13细胞中沉默QKI-5的细胞增殖和凋亡情况。采用Western blot方法检测SCC25细胞中过表达QKI-5和PCI13细胞中沉默QKI-5中细胞周期蛋白D1(Cyclin D1)、CDK4、P27、PARP-cleaved蛋白的表达情况。

结果

SCC15和SCC25细胞中QKI-5的蛋白和mRNA表达水平较低,而PCI13细胞中的QKI-5蛋白和mRNA表达水平较高。采用病毒介导的QKI-5过表达后,SCC25细胞中QKI-5的蛋白和mRNA水平显著增加。采用病毒介导的QKI-5沉默后,PCI13细胞中QKI-5的蛋白和mRNA水平显著减少。SCC25细胞中过表达QKI-5后,细胞增殖减少(P < 0.05),细胞多停留在G0/G1期,而凋亡增加,同时Cyclin D1和CDK4表达水平下调(P < 0.05),P27和PAPR-clevead蛋白表达水平上调(P < 0.05)。PCI13细胞中的QKI-5被沉默后,细胞增殖增加(P < 0.05),细胞多进入S期,而凋亡减少,同时Cyclin D1和CDK4表达水平上调(P < 0.05),P27和PARP-cleaved表达水平下调(P < 0.05)。

结论

QKI-5的表达可对人头颈鳞癌细胞增殖产生抑制作用。

Objective

To study the inhibition of QKI-5 on human head and neck cancer cell lines in vitro.

Methods

Western blot and immunohistochemistry were performed to detect the expression of QKI-5 in the human tongue squamous-cell carcinoma samples. Head and neck cancer cells line PCI13 and tongue squamous cell carcinoma cells lines SCC15 and SCC25 were cultured by the density of 5 × 104/ml. Western blot and RT-PCR were used to detect the expression levels of protein and mRNA of QKI-5 in the three cell lines. Using viral to mediate QKI-5 overexpressing in SCC25 cells and silencing in PCI13 cells. MTT method and flow cytometry assay were conducted to detect the proliferation and apoptosis of SCC25 cells with QKI-5 overexpressing and PCI13 cells with QKI-5 silencing. Western blot was used for the detection of cyclin D1, CDK4, P27, and PARP-cleaved in SCC25 cells with QKI-5 overexpressing and PCI13 cells with QKI-5 silencing.

Results

The RNA and protein expression levels of QKI-5 were relatively low in SCC15 and SCC25 cells, while in which the PCI13 cells was higher. Protein and RNA levels of QKI-5 increased significantly after viral mediated overexpression in SCC25 cells, and which were decreased significantly after virus mediated QKI-5 silencing in PCI13 cells. After overexpression of QKI-5 in SCC25 cells, the proliferation of SCC25 decreased, and most cells stayed in the G0/G1 phase, while cell apoptosis increased (P < 0.05) , the expression level of cyclin D1 and CDK4 decreased (P < 0.05) and the expression level of P27 and PAPR-cleaved increased (P < 0.05) . After QKI-5 silencing in PCI13 cells, the cell proliferation of PCI13 increased (P < 0.05) , most cells went into S phase, while apoptosis decreased, and the expression level increased in cyclin D1 and CDK4 (P < 0.05) , and decreased in P27 and PARP-cleaved (P < 0.05) .

Conclusions

QKI-5 could inhibit the proliferation of head and neck cancer cells.

图1 人头颈鳞癌中QKI-5蛋白的表达情况(SP染色)
图2 头颈鳞癌SCC15、SCC25和PCI13细胞株中QKI-5的表达情况
图3 SCC25细胞中过表达QKI-5和在PCI13细胞中沉默QKI-5后对QKI-5表达的影响
图4 QKI-5对头颈鳞癌细胞的细胞生长的影响
图5 QKI-5对头颈鳞癌细胞的细胞周期和凋亡的影响
图6 SCC25细胞中过表达QKI-5和在PCI13细胞中沉默QKI-5
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