转染环氧合酶-2反义寡核苷酸对骨肉瘤细胞侵袭性的抑制作用

目的 探讨环氧合酶-2(COX-2)反义寡核苷酸(ODNs)对人骨肉瘤细胞株OS-732侵袭性的抑制作用及其机制.方法 设计和合成COX-2反义寡核苷酸,体外转染骨肉瘤OS-732细胞,逆转录.聚合酶链反应(RT-PCR)及Western blot证实转染效果.改良Boyden-transwell方法观察肿瘤细胞侵袋抑制率,RT-PCR方法研究转染COX-2反义ODNs对OS-732细胞尿激酶型纤溶酶原激活物及其受体(uPA、uPAR)mRNA表达的影响.结果 转染COX-2反义ODNs的细胞COX-2核酸、蛋白表达水平显著降低.转染组细胞侵袭能力显著降低,呈浓度依赖性.转染组细胞uPA、uPAR的mRNA表达水平显著降低(P<0.01).结论 COX-2反义ODNs对OS-732细胞侵袭能力有明显抑制作用,其对uPA、uPAR表达的下调是主要作用机制.

The inhibitory effects of cyclooxygenase-2 antisense oligodeoxynucleotides on the invasion of osteosarcoma OS-732 cell line

Objective To investigate the inhibitory effects of cyclooxygenase-2 (COX-2) antisense oligodeoxynucleotides (anti-ODNs) on the invasion of osteosarcoma OS-732 cell line and the mechanism. Methods The COX-2 anti-ODNs were designed,synthesized and transfected into OS-732 cell line. The transfection effects were confrimed by RT-PCR and Western blot. The invasion inhibition ratio of OS732 cells was detected by modified Boyden-transwell techniques. The effects of COX-2 anti-ODNs on mRNA expression of uPA and uPAR were studied by RT-PCR. Results The expression of COX-2 mRNA and protein was decreased after transfecting COX-2 anti-ODNs into OS-732 cell line in a certain degree of dose-dependent manner. The expression of uPA and uPAR mRNA in transfected OS-732 cells were notably decreased by RT-PCR compared with control groups. Conclusion COX-2 anti-ODNs could remarkably inhibit the invasion ability of OS-732 cells probably by down-regulating the expression of uPA and uPAR.