A novel uPAg-KPI fusion protein inhibits the growth and invasion of human ovarian cancer cells in vitro.
A novel uPAg-KPI fusion protein inhibits the growth and invasion of human ovarian cancer cells in vitro.
复制标题
新型uPAg-KPI融合蛋白体外抑制人卵巢癌细胞的生长和侵袭
DOI：
10.3892/ijmm.2016.2540
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发表时间：
2016-05
影响因子：
5.4
通讯作者：
Cui MH
中科院分区：
文献类型：
--
作者：
Zhao LP;Xu TM;Kan MJ;Xiao YC;Cui MH
尿激酶型纤溶酶原激活物(UPA)通过破坏基底膜发挥作用，参与细胞的增殖、迁移和侵袭。这些作用是通过其生长因子结构域(GFD)与uPA受体(UPAR)结合而实现的。本研究评估了uPA-GFD和淀粉样蛋白β前体中存在的kpI结构域的融合蛋白uPAg-kpI的作用。使用卵巢癌细胞株SKOV-3，我们检测了细胞活力、迁移、侵袭和蛋白质表达。此外，我们使用Transwell分析检查了伤口愈合、迁移和侵袭。我们的数据显示，uPAg-KPI处理通过将肿瘤细胞阻滞在细胞周期的G1/G0期，以浓度和时间依赖的方式降低卵巢癌SKOV-3细胞的活力。作用48h后，uPAg-KPI的IC50值为0.5微克/微克L。在该浓度下，uPAg-KPI还抑制肿瘤细胞集落形成、伤口闭合以及细胞迁移和侵袭能力。蛋白质水平的蛋白质印迹分析表明，uPAg-KPI对细胞外信号调节蛋白1/ERK2和AKT的表达无明显影响，但对磷酸化的ERK1/ERK2和AKT的表达有抑制作用。因此，我们认为这种新型的uPAg-KPI融合蛋白通过调节ERK和AKT信号通路，在体外降低了人卵巢癌SKOV-3细胞的存活率、克隆形成、伤口愈合和侵袭能力。使用其他细胞系的进一步研究将证实这些发现。
Urokinase-type plasminogen activator (uPA) acts by breaking down the basement membrane and is involved in cell proliferation, migration and invasion. These actions are mediated by binding to the uPA receptor (uPAR) via its growth factor domain (GFD). The present study evaluated the effects of uPAg-KPI, a fusion protein of uPA-GFD and a kunitz protease inhibitor (KPI) domain that is present in the amyloid β-protein precursor. Using SKOV-3 cells, an ovarian cancer cell line, we examined cell viability, migration, invasion and also protein expression. Furthermore, we examined wound healing, and migration and invasion using a Transwell assay. Our data showed that uPAg-KPI treatment reduced the viability of ovarian cancer SKOV-3 cells in both a concentration and time-dependent manner by arresting tumor cells at G1/G0 phase of the cell cycle. The IC50 of uPAg-KPI was 0.5 µg/µl after 48 h treatment. At this concentration, uPAg-KPI also inhibited tumor cell colony formation, wound closure, as well as cell migration and invasion capacity. At the protein level, western blot analysis demonstrated that uPAg-KPI exerted no significant effect on the expression of total extracellular signal-regulated kinase (ERK)1/ERK2 and AKT, whereas it suppressed levels of phosphorylated ERK1/ERK2 and AKT. Thus, we suggest that this novel uPAg-KPI fusion protein reduced cell viability, colony formation, wound healing and the invasive ability of human ovarian cancer SKOV-3 cells in vitro by regulating ERK and AKT signaling. Further studies using other cell lines will confirm these findings.