Sprouty 1蛋白的表达与卵巢癌细胞的生长、增殖、迁移和侵袭呈负相关。
The expression of the Sprouty 1 protein inversely correlates with growth, proliferation, migration and invasion of ovarian cancer cells.
作者信息
Masoumi-Moghaddam Samar, Amini Afshin, Ehteda Anahid, Wei Ai-Qun, Morris David Lawson
机构信息
Department of Surgery, St George Hospital, The University of New South Wales, Gray Street, Kogarah, Sydney NSW 2217, Australia.
Department of Orthopedic Surgery, St. George Hospital, The University of New South Wales, Gray Street, Kogarah, Sydney NSW 2217, Australia.
出版信息
J Ovarian Res. 2014 Jun 8;7:61. doi: 10.1186/1757-2215-7-61. eCollection 2014.
BACKGROUND
Our recent study on a panel of human ovarian cancer cells revealed that SKOV-3 cells barely express the Sprouty isoform 1 (Spry1) while 1A9 cells maintain it at a level similar to normal ovarian cells. Here we investigated the functional outcomes of induced alterations in the expression of Spry1 in the two cell lines in vitro.
METHODS
Using the Spry1 specific plasmid and siRNA, the expression of Spry1 was induced and conversely silenced in SKOV-3 and 1A9 cells, respectively. The functional outcome was investigated by means of proliferation, MTT, scratch-wound, migration and invasion assays and selection of the stable clones. Mechanism of the effect was explored by Western blot.
RESULTS
In the Spry1-transfected SKOV-3 cells, a significant reduction in growth and proliferation was evident. Stable clones of the Spry1-transfected SKOV-3 were almost undetectable after day 14. The number of migrated and invaded cells and the percentage of the scratch closure were significantly lower in the Spry1-transfected group. Spry1 silencing in 1A9 cells, on the other hand, led to a significant increase in cell growth and proliferation. The number of migrated and invaded cells and the percentage of the scratch closure significantly increased in Spry1-silenced 1A9 group. Mechanistically, overexpression of Bax, activation of caspases 3, 7, 8 and 9, cleavage of PARP and attenuation of Bcl-2 and Bcl-xl were observed along with reduced activation of Erk and Akt and increased amount and activity of PTEN in the Spry1-transfected SKOV-3 cells.
CONCLUSIONS
Here, we report the inverse correlation between the expression of Spry1 and growth, proliferation, invasion and migration of ovarian cancer cells.
背景
我们最近对一组人卵巢癌细胞的研究表明，SKOV-3细胞几乎不表达Sprouty异构体1（Spry1），而1A9细胞维持其表达水平与正常卵巢细胞相似。在此，我们研究了体外诱导这两种细胞系中Spry1表达改变的功能结果。
方法
分别使用Spry1特异性质粒和小干扰RNA（siRNA），在SKOV-3和1A9细胞中诱导Spry1表达并使其反向沉默。通过增殖、MTT、划痕、迁移和侵袭实验以及稳定克隆的筛选来研究功能结果。通过蛋白质免疫印迹法探索作用机制。
结果
在转染Spry1的SKOV-3细胞中，生长和增殖明显降低。转染Spry1的SKOV-3细胞在第14天后几乎检测不到稳定克隆。转染Spry1的组中迁移和侵袭的细胞数量以及划痕闭合百分比显著降低。另一方面，1A9细胞中Spry1沉默导致细胞生长和增殖显著增加。Spry1沉默的1A9组中迁移和侵袭的细胞数量以及划痕闭合百分比显著增加。机制上，在转染Spry1的SKOV-3细胞中观察到Bax过表达、半胱天冬酶3、7、8和9激活、PARP裂解以及Bcl-2和Bcl-xl减弱，同时Erk和Akt激活减少，PTEN量和活性增加。
结论
在此，我们报道Spry1表达与卵巢癌细胞生长、增殖、侵袭和迁移之间呈负相关。
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