摘要
目的探讨外源性骨桥蛋白(OPN)对卵巢癌细胞(SKOV3)生长及迁移侵袭能力的影响。方法选用SKOV3及人正常卵巢上皮细胞(IOSE80)。应用Elisa及蛋白印迹方法检测OPN含量。CCK8方法测定细胞增殖。划痕实验及小室穿梭实验评价细胞迁移侵袭能力。蛋白印迹的方法检测细胞信号通路活化及蛋白表达量。结果 OPN的分泌量及表达量在SKOV3细胞中均高于IOSE80(P均<0.05)。OPN浓度分别为20、50、80、110 ng/m L时,均促进了SKOV3的增殖(P均<0.05)。空白对照组及OPN(50 ng/m L)组细胞运动面积分别为(26.1±0.9)%和(42.9±1.5)%(P<0.05)。空白对照组与OPN(50 ng/m L)组跨膜细胞数量分别为(165±7.63)个/孔和(333.3±8.8)个/孔(P<0.05)。OPN(50 ng/m L)干预15 min明显上调磷酸化Akt及MMP2表达量。应用PI3K抑制剂预处理后,OPN(50 ng/m L)干预对磷酸化Akt及MMP2表达量没有明显影响。结论 SKOV3比IOSE80表达分泌更多OPN。外源性OPN促进了SKOV3的增殖、迁移侵袭能力。OPN激活PI3K通路上调MMP2是可能机制之一。
Objective To investigate the impact of osteopontin （OPN） on proliferation and invasion of ovarian cancer Cell line （SKOV3）. Methods Routinely cultured SKOV3 cells were treated with OPN for certain times. And cell num- bers were measured using CCK8. The cell migration and invasion were assessed by wound healing test and transwell chamber assay, respectively. Elisa and western blotting were used to detect the protein expressions of p-Akt, Akt and MMP2. Results Compared with IOSE80 ceils, SKOV3 secreted and expressed more OPN （P〈0.05）. Compared with control group, 20, 50, 80, 110 ng/mL OPN promote proliferation of SKOV3 cells （P〈0.05）. SKOV3 cells treated with 50 ng/mL OPN showed increased ability of migration and invasion （both P〈0.05）. 50 ng/mL OPN treated for 15 minutes significantly increased the phosphorylation of Akt and the protein expressions of MMP2 of SKOV3. Specifi- cally blocked the phosphorylation of PI3K inhibited the phosphorylation of Akt and the protein expressions of MMP2 when treated with OPN. Conclusion OPN could promote the migration and invasion of SKOV3 cells possibly by regu- lating the phosphorylation of PI3K and the expression of MMP2.
出处
《中国现代医生》
2014年第36期1-4,8,共5页
China Modern Doctor
基金
黑龙江省留学归国人员科学基金项目(LC2011C26)
关键词
卵巢癌
骨桥蛋白
基质金属蛋白酶
Ovarian cancer
Osteopontin
Matrix metalloproteinase
