环氧化酶-2反义核酸对膀胱癌细胞株裸鼠成瘤的影响

Inhibitive effect of the Cyclooxygenase-2 antisense nucleic acid on human bladder cancer cell xenografts in nude mice

目的探讨环氧化酶-2反义核酸对膀胱癌细胞株裸鼠成瘤的影响。方法应用脂质体介导获得了稳定表达人环氧化酶-2反义核酸、空载体的膀胱癌细胞T24-AS、T24-P。BALB/c裸鼠27只分为3组,每只皮下分别接种0.2mLT24-AS、T24-P、T24膀胱癌细胞(2.5×107/mL)。30d后处死裸鼠,取瘤块称重、测量肿瘤体积。免疫组化、半定量RT-PCR、Wester blot检测移植瘤COX-2表达。结果裸鼠成瘤试验T24-P、T24组中,细胞接种裸鼠后第5天可见肿瘤长出,T24-AS组接种后第7天可见肿瘤长出。T24、T24-AS、T24-P组移植瘤重分别为(965.5±84.9)mg、(66.7±45.1)mg、(1 023.5±40.2)mg。免疫组化染色、RTPCR、Western blot结果均显示T24、T24-P组COX-2表达明显,而T24-AS组少量表达。结论环氧化酶-2反义核酸在实验中表现出良好的抗肿瘤特性。

Objective To establish the nude mice xenografts models of the bladder cancer cell T24 transfected with the antisense Cyclooxygenase-2 （COX-2） RNA and explore the relationship between COX-2 gene and the growth inhibition effects of mice xenografts. Method Using lipofect AMINETM2000 reagent, to transfect the antisense eukaryotic expression vector pcDNA3.1/hCOX-2 and control plasmid pcDNA3.1 into the bladder cancer cells T24 respectively to get the T24-AS and the contrast （T24-P） bladder cells. The 27 athymic BALB/c mice were divided equally into three groups. Each 9 of them was injec- ted into 5 × 10^6 T24 bladder cancer cells （T24 or T24-AS or T24-P） separately. After 30 days, the nude mice were killed and the weight and volume of the xenografts were recorded. Histological hematoxylin-osin staining and immunocytochemical SABC technique were employed to detect the histopathological changes. To detect expression of COX-2 in human bladder cancer cell xenografts by RT-PCR and western blot. Results On the day 5 after transplantation, xenografts was found in nude mice in- jected T24 and T24-P cells. Two days later, in the rest of the mices xenografts was found too. The weights of the xenografts （T24 or T24-AS or T24-P） were （965.5±84.9）mg, （66.7±45.1）mg, （1 023.5±40.2）mg separately. The expression level of COX-2 decreased and the weight and volume of the xenografts decreased more in the groups of T24-AS. Conclusions The Cyclooxygenase-2 antisense nucleic acid could inhibit the xenografts growth in nude mice.