Xia Yong-Hua, Li Min, Fu Dan-Dan, Xu Su-Ling, Li Zhan-Guo, Liu Dong, Tian Zhong-Wei
Department of Dermatovenereology, the First Affiliated Hospital of Xinxiang Medical University, Xinxiang, Henan, China E-mail :
Asian Pac J Cancer Prev. 2013;14(11):6245-8. doi: 10.7314/apjcp.2013.14.11.6245.
To study effects of down-regulation of pituitary tumor-transforming gene (PTTG) on proliferation and metastasis ability of the SCL-1 cutaneous squamous cell carcinoma (CSCC) cell line and explore related mechanisms.
SCL-1 cells were divided into 3 groups (untreated, siRNA control and PTTG siRNA). Cell proliferation assays were performed using a CCK-8 kit and proliferation and metastasis ability were analyzed using Boyden chambers. In addition, expression of MMP-2 and MMP-9 was detected by r-time qPCR and Western blotting.
Down-regulation of PTTG could markedly inhibit cell proliferation in SCL-1 cells, compared to untreated and control siRNA groups (P < 0.05). Real-time qPCR demonstrated that expression levels of PTTG, MMP-2 and MMP-9 in the PTTG siRNA group were 0.8%, 23.2% and 21.3% of untreated levels. Western blotting revealed that expression of PTTG, MMP-2 and MMP-9 proteins in the PTTG siRNA group was obviously down-regulated. The numbers of migrating cells (51.38 ± 4.71) in the PTTG siRNA group was obviously lower than that in untreated group (131.33 ± 6.12) and the control siRNA group (127.72 ± 5.20) (P < 0.05), suggesting that decrease of proliferation and metastasis ability mediated by PTTG knock-down may be closely correlated with down-regulation of MMP-2 and MMP-9 expression.
Inhibition of PTTG expression may be a new target for therapy of CSCC.
研究垂体肿瘤转化基因(PTTG)下调对SCL-1皮肤鳞状细胞癌(CSCC)细胞系增殖和转移能力的影响,并探讨相关机制。
将SCL-1细胞分为3组(未处理组、siRNA对照组和PTTG siRNA组)。使用CCK-8试剂盒进行细胞增殖检测,并使用Boyden小室分析增殖和转移能力。此外,通过实时定量PCR和蛋白质印迹法检测MMP-2和MMP-9的表达。
与未处理组和对照siRNA组相比,PTTG下调可显著抑制SCL-1细胞的增殖(P<0.05)。实时定量PCR显示,PTTG siRNA组中PTTG、MMP-2和MMP-9的表达水平分别为未处理组的0.8%、23.2%和21.3%。蛋白质印迹法显示,PTTG siRNA组中PTTG、MMP-2和MMP-9蛋白的表达明显下调。PTTG siRNA组中迁移细胞的数量(51.38±4.71)明显低于未处理组(131.33±6.12)和对照siRNA组('127.72±5.20)(P<0.05),提示PTTG敲低介导的增殖和转移能力降低可能与MMP-2和MMP-9表达下调密切相关。
抑制PTTG表达可能是CSCC治疗的新靶点。