中华皮肤科杂志 ›› 2010, Vol. 43 ›› Issue (3): 174-177.

• 论著 • 上一篇    下一篇

下调垂体瘤转化基因对皮肤鳞状细胞癌细胞SCL-1细胞增殖及迁移能力的影响

刘冬1,夏永华1,李敏河南2,李占国3,付丹丹3,田中伟2   

  1. 1. 新乡医学院第一附属医院皮肤性病科
    2. 河南省新乡医学院第一附属医院皮肤科
    3.
  • 收稿日期:2009-05-22 修回日期:2009-07-13 出版日期:2010-03-15 发布日期:2012-03-31
  • 通讯作者: 刘冬 E-mail:liudong0064@163.com

Effect of down-regulation of pituitary tumor-transforming gene （PTTG） on the proliferation and migration of cutaneous squamous cell carcinoma cell line SCL-1

  • Received:2009-05-22 Revised:2009-07-13 Online:2010-03-15 Published:2012-03-31

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摘要： 目的 研究下调垂体瘤转化基因（PTTG）的表达对皮肤鳞状细胞癌（鳞癌）细胞SCL-1细胞增殖及浸润转移能力的影响，探讨其相关的分子机制。方法 将SCL-1细胞分为三组（未处理组、转染对照siRNA组及转染PTTG siRNA组），首先利用CCK-8试剂盒研究下调PTTG对SCL-1细胞增殖能力的影响，并利用Boyden chamber研究三组细胞的浸润转移能力，通过实时荧光定量PCR和Western印迹方法检测MMP-2和MMP-9在三组细胞中的表达。结果 与未处理组和转染对照siRNA组相比，转染PTTG siRNA后能明显抑制SCL-1细胞增殖（P < 0.05）。此外，实时荧光定量PCR结果显示，转染PTTG siRNA后，PTTG、MMP-2和MMP-9的表达均明显下降，表达水平分别是对照组的0.8%、23.2%和21.3%。Western印迹结果表明，转染PTTG siRNA后，PTTG、MMP-2和MMP-9的蛋白表达水平也明显下降（P < 0.05）。转染PTTG siRNA组的细胞穿膜数（51.38 ± 4.71）明显低于未处理组（131.33 ± 6.12）及转染siRNA对照组（127.72 ± 5.20）（P < 0.05）。结论 抑制PTTG的表达能显著抑制SCL-1细胞增殖及迁移，且可下调MMP-2和MMP-9表达。

关键词: 癌，鳞状细胞, 垂体瘤转化基因, RNA，小分子干扰, 细胞增殖, 细胞系，肿瘤, 肿瘤浸润

Abstract: Objective To study the effect of down-regulation of PTTG on the proliferation and migration of cutaneous squamous cell carcinoma cell line SCL-1 and its related mechanism. Methods SCL-1 cells were transfected with control siRNA or PTTG-targeting siRNA （PTTG-siRNA）, or remained untransfected. After additional culture, the proliferation of SCL-1 cells as observed with cell counting kit-8 （CCK-8）, and cell migration with Boyden chamber. Real-time PCR and Western blot were performed to detect the expression of matrix metalloproteinase 2 （MMP-2）, MMP-9 and PTTG. Results The proliferation of SCL-1 cells transfected with PTTG-siRNA was markedly deccelarated in comparision with that of untransfected cells and those transfected with control siRNA （both P < 0.05）. Real-time PCR and Western blot disclosed a significant decrease in the mRNA and protein expression of MMP-2, MMP-9 and PTTG in PTTG-siRNA-transfected SCL-1 cells compared with the other two groups of cells. As real-time PCR showed, the expressions of MMP-2, MMP-9 and PTTG in PTTG-siRNA-transfected SCL-1 cells were 0.8%, 23.2% and 21.3% of those in untransfected cells, respectively. Further more, the number of SCL-1 cells migrating through microporous membrane in the Boyden chamber was significantly smaller in PTTG-siRNA-transfected group than in untransfected group and control siRNA-transfected group （51.38 ± 4.71 vs 131.33 ± 6.12 and 127.72 ± 5.20, both P < 0.05）. Conclusion The down-regulation of PTTG may deccelarate the proliferation and migration of SCL-1 cells and inhibit the expression of MMP-2 and MMP-9 in SCL-1 cells.

Key words: Pituitary tumor-transforming gene, cell proliferation