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

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

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fyITrGsiRNA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Boydenchamber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PTYGsiRNA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PTTGsiRNA后，PTYG、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PTrGsiRNA后，P1TG、MMP-2和MMP-9的蛋白表达水平也明显下降（P〈0．05）。转染PTTGsiRNA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。

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 migra- tion 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 ceils, respectively. Further more, the number of SCL-1 cells migrating through microporous membrane in the Boyden chamber was significantly smaller in PTI＇G-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 ceils and inhibit the expression of MMP-2 and MMP-9 in SCL-1 cells.