低浓度Celecoxib对卵巢癌细胞SKOV3侵袭的抑制作用的研究  

Low-concentration Celecoxib inhibits the invasiveness of human ovarian cancer cell line SKOV3

作　　者：杨红[1] 邹伟[1] 辛晓燕[1] 

机构地区：[1]第四军医大学西医京院妇产科,西安710032

出　　处：《中国药物与临床》2008年第9期684-688,753,共6页Chinese Remedies & Clinics

基　　金：国家自然科学基金资助项目(30772305);西京医院创新基金资助项目(XJCX06M25)

摘　　要：目的探讨低浓度环氧化酶-2(COX-2)选择性抑制剂Celecoxib对人卵巢癌细胞系SKOV3侵袭的影响及其相关机制。方法10μmol/L Celecoxib作用SKOV3细胞24h后,细胞外基质黏附试验检测细胞黏附能力,Transwell侵袭小室测定细胞的侵袭力,划痕试验检测细胞的迁移能力,明胶酶谱法(Zymography)检测细胞基质金属蛋白酶(MMP)-2、MMP-9活性,Western blot检测MMP-2、MMP-9和E-cadherin的表达;同时利用RNAi技术特异性沉默SKOV3细胞中COX-2的表达,观察上述指标的变化情况。结果10μmol/L Celecoxib作用SKOV3细胞24h后,细胞外基质黏附试验结果表明:SKOV3细胞黏附于matrigel的细胞数明显增加(P<0.05);Transwell侵袭小室实验结果表明:SKOV3细胞穿膜数明显减少(P<0.05),划痕试验结果表明:SKOV3细胞迁移到损伤区的细胞数明显减少(P<0.05)。进一步探讨机制,Zymography结果显示Celecoxib作用SKOV3细胞后,MMP-2、MMP-9的活性降低,Western blot结果显示MMP-2、MMP-9的表达量降低,而E-cadherin的表达量增强。而经特异性沉默COX-2表达的SKOV3/COX-2i细胞未出现上述分子的变化。结论10μmol/L Celecoxib能够增强SKOV3的黏附力,抑制其侵袭和转移能力,其机制可能与增加E-cadherin的表达,抑制MMP-2、MMP-9的活性和表达有关,而这一过程可能是COX-2非依赖的。Objective To investigate the protection of Celecoxib, a cyclooxygenase-2 selective inhibitor, against the invasiveness of human ovarian cancer cell line SKOV3 and its potential mechanism. Methods Following 24-hour incubation with 10 μmol/L Celecoxib, the SKOV3 cells were detected for cell adhesion, invasion and migration using cell adhesion assay, transwell chamber and scratch assay, The activities of MMP-2 and MMP-9 were examined with Zymography, and the protein expressions of COX-2, MMP-2, MMP-9 and E-cadherin were analyzed with Western blotting. The COX-2 siRNA expression vector was transfected into SKOV3 cells to investigate the potential role of COX-2 in the changes caused by Celecoxib. Results After SKOV3 cells were treated with 10 μmol/L Celecoxib for 24 hours, cell adhesive assay, Transwell chamber assay and scratch assay indicated enhanced adhesive ability and significantly inhibited invasiveness and migratory ability （all P〈0.05）. Zymography results showed reduced activities of MMP-2 and MMP-9, whereas Western blotting showed decreased protein expressions of MMP-2, MMP-9 and increased protein expressions of E-cadherin, But the same changes were not observed in SKOV3/COX-2i cells transfected with COX-2 siRNA expression vector. Conclusion 10 μmol/L Celecoxib was demonstrated to enhance the adhesive ability and suppress the invasive and migratory abilities of SKOV3 cells. The underlying mechanism may be associated with down-regulation of MMP-2/MMP-9 and up-regulation of E-cadherin, which may be independent of COX-2 expression.

关 键 词：卵巢肿瘤 肿瘤浸润 环氧化酶-2选择性抑制剂 

分 类 号：R737.31[医药卫生—肿瘤]