人卵巢癌相关抗原66对人脑胶质瘤细胞U251迁移、侵袭的影响及其机制

Effect and the mechanism of the ovarian associated antigen 66 on cell migration and invasion of human brain glioma U251 cells

目的 探讨人卵巢癌相关抗原66（OVA66）对人脑胶质瘤细胞U251迁移、侵袭的影响及其机制.方法 运用Control-小干扰RNA （siRNA）、OVA66-siRNA分别转染人脑胶质瘤细胞U251,Western blot检测OVA66蛋白表达及基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶-9（MMP-9）,细胞划痕实验及Transwell迁移、侵袭实验检测细胞的迁移、侵袭能力.结果 与Control-siRNA组比较OVA66-siRNA组人脑胶质瘤细胞U251蛋白表达量降低63.2％,MMP-2、MMP-9的蛋白表达量分别降低58.0％和27.1％,差异有统计学意义（P＜0.01）.细胞划痕实验结果显示OVA66-siRNA组较Control-siRNA组细胞的迁移能力明显降低（P＜0.05）.Transwell迁移、侵袭实验结果如下：Control-siRNA组迁移细胞为（153.0±5.2）个,OVA66-siRNA组迁移细胞为（80.0±3.6）个,OVA66-siRNA组迁移细胞数目明显减少（P＜0.01）;Control-siRNA组侵袭细胞为（142.0±8.5）个,OVA66-siRNA组侵袭细胞为（51.0±8.1）个,OVA66-siRNA组侵袭细胞数目明显减少（P＜0.01）.结论 OVA66-siRNA靶向干扰了OVA66的表达,同时下调MMP-2、MMP-9的表达.下调OVA66的表达水平能够降低人脑胶质瘤细胞U251迁移、侵袭的能力。

Objective To investigate the effect and the mechanism of ovarian associated antigen 66 （OVA66） on cell migration and invasion of human brain glioma U251 cells.Methods Control-small interference RNA （siRNA） and OVA66-siRNA were used respectively to transfect human brain glioma U251 cells.The expression levels of OVA66 and Matrix metallopeptidase-2 （MMP-2）,Matrix metallopeptidase-9 （MMP-9） proteins in human brain glioma U251 cells were both detected after OVA66 knockdown by using Western blotting.The migration and invasion ability of U251 cells were respectively measured by wound-healing assay and transwell migration and invasion assays.Results As compared with control-siRNA group,the expression of OVA66 was decreased by 63.2% in OVA66-siRNA group and the expression levels of MMP-2 and MMP-9 were both drastically decreased （P 〈0.01）.The migration ability of U251 cells was decreased in wound-healing assay after OVA66 knockdown （P 〈 0.05）.The number of migrated cells in group Control-siRNA and OVA66-siRNA were 153.0 ± 5.2 and 80.0 ± 3.6,respectively.The number of invaded cells in group Control-siRNA and OVA66-siRNA were 142.0 ± 8.5 and 51.0 ± 8.1,respectively.These indicated that cell migration and invasion ability of transfected cells were decreased （P 〈 0.01）.Conclusion OVA66-targeted siRNA interferes with the expression levels of OVA66,and reduced the expression levels of MMP-2 and MMP-9.OVA66 acts as a tumor promotion that promotes the migration and invasion of glioma U251 cells.